Silk performance apparel and products and methods of preparing the same

ABSTRACT

Silk performance apparel and methods of preparing the same are disclosed herein. In some embodiments, silk performance apparel includes textiles, fabrics, consumer products, and other materials that are coated with aqueous solutions of pure silk fibroin-based protein fragments. In some embodiments, coated apparel products exhibit surprisingly improved moisture management properties and increased resistance to microbial growth.

RELATED APPLICATIONS

The present application is a continuation of U.S. Nonprovisional Utilityapplication Ser. No. 14/958,565, filed Dec. 3, 2015, which is acontinuation of international application no. PCT/US2015/063545 filedDec. 2, 2015, which application claims the benefit of U.S. ProvisionalApplication No. 62/086,297 filed Dec. 2, 2014 and U.S. ProvisionalApplication No. 62/192,477 filed Jul. 14, 2015 and U.S. ProvisionalApplication No. 62/245,221 filed Oct. 22, 2015. The contents of each ofthese applications are incorporated herein by reference in theirentireties.

FIELD OF THE INVENTION

In some embodiments, the present invention relates to silk performanceapparel and products, such as fabrics coated with pure silkfibroin-based proteins or protein fragments.

BACKGROUND OF THE INVENTION

Silk is a natural polymer produced by a variety of insects and spiders,and comprises a filament core protein, silk fibroin, and a glue-likecoating consisting of a non-filamentous protein, sericin. Silk fibersare light weight, breathable, and hypoallergenic. Silk is comfortablewhen worn next to the skin and insulates very well; keeping the wearerwarm in cold temperatures and is cooler than many other fabrics in warmtemperatures.

SUMMARY OF THE INVENTION

Silk performance apparel and methods of preparing the same are disclosedherein. According to aspects illustrated herein, the present disclosurerelates to a product, including, but not limited to, apparel, padding,shoes, gloves, luggage, furs, jewelry and bags, configured to be worn orcarried on the body, that is at least partially surface treated with anaqueous solution of pure silk fibroin-based protein fragments of thepresent disclosure so as to result in a silk coating on the product. Inan embodiment, the product is manufactured from a textile material. Inan embodiment, the product is manufactured from a non-textile material.In an embodiment, desired additives can be added to an aqueous solutionof pure silk fibroin-based protein fragments of the present disclosureso as to result in a silk coating having desired additives.

According to aspects illustrated herein, an aqueous solution of puresilk fibroin-based protein fragments of the present disclosure isavailable in a spray can for spraying on a product, including, but notlimited to, apparel, padding, shoes, gloves, luggage, furs, jewelry andbags, or for directly spraying on the body of a consumer, to impartdesired properties to the product. In an embodiment, the product ismanufactured from a textile material. In an embodiment, the product ismanufactured from a non-textile material. In an embodiment, desiredadditives can be added to an aqueous solution of pure silk fibroin-basedprotein fragments of the present disclosure so as to result in a silkcoating having desired additives.

In an embodiment, a textile comprising a silk coating of the presentdisclosure is sold to a consumer. In an embodiment, a textile of thepresent disclosure is used in constructing action sportswear apparel. Inan embodiment, a textile of the present disclosure is used inconstructing fitness apparel. In an embodiment, a textile of the presentdisclosure is used in constructing performance apparel. In anembodiment, a textile of the present disclosure is used in constructinggolf apparel. In an embodiment, a textile of the present disclosure isused in constructing lingerie. In an embodiment, a silk coating of thepresent disclosure is positioned on the underlining of actionsportswear/apparel. In an embodiment, a silk coating of the presentdisclosure is positioned on the shell, the lining, or the interlining ofaction sportswear/apparel. In an embodiment, action sportswear/apparelis partially made from a silk coated textile of the present disclosureand partially made from an uncoated textile. In an embodiment, actionsportswear/apparel partially made from a silk coated textile andpartially made from an uncoated textile combines an uncoated inertsynthetic material with a silk coated inert synthetic material. Examplesof inert synthetic material include, but are not limited to, polyester,polyamide, polyaramid, polytetrafluorethylene, polyethylene,polypropylene, polyurethane, silicone, mixtures of polyurethane andpolyethylenglycol, ultrahigh molecular weight polyethylene,high-performance polyethylene, nylon, LYCRA (polyester-polyurethanecopolymer, also known as SPANDEX and elastomer), and mixtures thereof.In an embodiment, action sportswear/apparel partially made from a silkcoated textile and partially made from an uncoated textile combines anelastomeric material at least partially covered with a silk coating ofthe present disclosure. In an embodiment, the percentage of silk toelastomeric material can be varied to achieve desired shrink or wrinkleresistant properties and desired moisture content against the skinsurface. In an embodiment, a silk coating of the present disclosure ispositioned on an internal layer of a shoe (textile or non-textilebased). In an embodiment, a silk coating of the present disclosurepositioned on an internal layer of a shoe helps maintain optimal feetmicroenvironment, such as temperature and humidity while reducing anyexcessive perspiration.

In an embodiment, a silk coating of the present disclosure is visible.In an embodiment, a silk coating of the present disclosure istransparent. In an embodiment, a silk coating of the present disclosurepositioned on action sportswear/apparel helps control skin temperatureof a person wearing the apparel. In an embodiment, a silk coating of thepresent disclosure positioned on action sportswear/apparel helps controlfluid transfer away from the skin of a person wearing the apparel. In anembodiment, a silk coating of the present disclosure positioned onaction sportswear/apparel has a soft feel against the skin decreasingabrasions from fabric on the skin. In an embodiment, a silk coating ofthe present disclosure positioned on a textile has properties thatconfer at least one of wrinkle resistance, shrinkage resistance, ormachine washability to the textile. In an embodiment, a silk coatedtextile of the present disclosure is 100% machine washable and drycleanable. In an embodiment, a silk coated textile of the presentdisclosure is 100% waterproof. In an embodiment, a silk coated textileof the present disclosure is wrinkle resistant. In an embodiment, a silkcoated textile of the present disclosure is shrink resistant. In anembodiment, a silk coated fabric improves the health of the skin. In anembodiment, healthy skin can be determined by visibly seeing an evenskin tone. In an embodiment, healthy skin can be determined by visiblyseeing a smooth, glowing complexion. In an embodiment, a silk coatedfabric decreases irritation of the skin. In an embodiment, a decrease inirritation of the skin can result in a decrease in skin bumps or sores.In an embodiment, a decrease in irritation of the skin can result in adecrease in scaly or red skin. In an embodiment, a decrease inirritation of the skin can result in a decrease in itchiness or burning.In an embodiment, a silk coated fabric decreases inflammation of theskin. In an embodiment, a silk coated textile of the present disclosurehas the qualities of being waterproof, breathable, and elastic andpossess a number of other qualities which are highly desirable in actionsportswear. In an embodiment, a silk coated textile of the presentdisclosure manufactured from a silk fabric of the present disclosurefurther includes LYCRA brand spandex fibers (polyester-polyurethanecopolymer).

In an embodiment, a textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure is a breathable fabric. In an embodiment, a textile at leastpartially coated with an aqueous solution of pure silk fibroin-basedprotein fragments of the present disclosure is a water-resistant fabric.In an embodiment, a textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure is a shrink-resistant fabric. In an embodiment, a textile atleast partially coated with an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure is amachine-washable fabric. In an embodiment, a textile at least partiallycoated with an aqueous solution of pure silk fibroin-based proteinfragments of the present disclosure is a wrinkle resistant fabric. In anembodiment, textile at least partially coated with an aqueous solutionof pure silk fibroin-based protein fragments of the present disclosureprovides moisture and vitamins to the skin.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has an accumulative one-way transport index of greater than140. In an embodiment, the textile at least partially coated with anaqueous solution of pure silk fibroin-based protein fragments of thepresent disclosure has an accumulative one-way transport index ofgreater than 120. In an embodiment, the textile at least partiallycoated with an aqueous solution of pure silk fibroin-based proteinfragments of the present disclosure has an accumulative one-waytransport index of greater than 100. In an embodiment, the textile atleast partially coated with an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure has anaccumulative one-way transport index of greater than 80.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has an overall moisture management capability of greater than0.4. In an embodiment, the textile at least partially coated with anaqueous solution of pure silk fibroin-based protein fragments of thepresent disclosure has an overall moisture management capability ofgreater than 0.35. In an embodiment, the textile at least partiallycoated with an aqueous solution of pure silk fibroin-based proteinfragments of the present disclosure has an overall moisture managementcapability of greater than 0.3. In an embodiment, the textile at leastpartially coated with an aqueous solution of pure silk fibroin-basedprotein fragments of the present disclosure has an overall moisturemanagement capability of greater than 0.25.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a wetting time of at least 3 seconds. In an embodiment,the textile at least partially coated with an aqueous solution of puresilk fibroin-based protein fragments of the present disclosure has awetting time of at least 2.5 seconds. In an embodiment, the textile atleast partially coated with an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure has a wettingtime of at least 2 seconds. In an embodiment, the textile at leastpartially coated with an aqueous solution of pure silk fibroin-basedprotein fragments of the present disclosure has a wetting time of atleast 1.5 seconds.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a top absorption time of at least 50 seconds. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a top absorption time of at least 40 seconds. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a top absorption time of at least 30 seconds.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a bottom absorption time of at least 80 seconds. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a bottom absorption time of at least 70 seconds. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a bottom absorption time of at least 60 seconds. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a bottom absorption time of at least 50 seconds. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a bottom absorption time of at least 40 seconds.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a spreading speed of at least 1.6 mm/second. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a spreading speed of at least 1.4 mm/second. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a spreading speed of at least 1.2 mm/second. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a spreading speed of at least 1.0 mm/second. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure has a spreading speed of at least 0.8 mm/second.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 2000% microbial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 1000% microbial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 500% microbial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 400% microbial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 300% microbial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 200% microbial growth over 24 hours.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 2000% bacterial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 1000% bacterial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 500% bacterial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 400% bacterial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 300% bacterial growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 200% bacterial growth over 24 hours.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 2000% fungal growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 1000% fungal growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 500% fungal growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 400% fungal growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 300% fungal growth over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 200% fungal growth over 24 hours.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 2000% growth of Staphylococcus aureus over 24hours. In an embodiment, the textile at least partially coated with anaqueous solution of pure silk fibroin-based protein fragments of thepresent disclosure shows less than 1000% growth of Staphylococcus aureusover 24 hours. In an embodiment, the textile at least partially coatedwith an aqueous solution of pure silk fibroin-based protein fragments ofthe present disclosure shows less than 500% growth of Staphylococcusaureus over 24 hours. In an embodiment, the textile at least partiallycoated with an aqueous solution of pure silk fibroin-based proteinfragments of the present disclosure shows less than 400% growth ofStaphylococcus aureus over 24 hours. In an embodiment, the textile atleast partially coated with an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure shows lessthan 300% growth of Staphylococcus aureus over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 200% growth of Staphylococcus aureus over 24hours.

In an embodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 2000% growth of Klebsiella pneumoniae over 24hours. In an embodiment, the textile at least partially coated with anaqueous solution of pure silk fibroin-based protein fragments of thepresent disclosure shows less than 1000% growth of Klebsiella pneumoniaeover 24 hours. In an embodiment, the textile at least partially coatedwith an aqueous solution of pure silk fibroin-based protein fragments ofthe present disclosure shows less than 500% growth of Klebsiellapneumoniae over 24 hours. In an embodiment, the textile at leastpartially coated with an aqueous solution of pure silk fibroin-basedprotein fragments of the present disclosure shows less than 400% growthof Klebsiella pneumoniae over 24 hours. In an embodiment, the textile atleast partially coated with an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure shows lessthan 300% growth of Klebsiella pneumoniae over 24 hours. In anembodiment, the textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure shows less than 200% growth of Klebsiella pneumoniae over 24hours.

In an embodiment, an aqueous solution of pure silk fibroin-based proteinfragments of the present disclosure is used to coat a textile. In anembodiment, the concentration of silk in the solution ranges from about0.1% to about 20.0%. In an embodiment, the concentration of silk in thesolution ranges from about 0.1% to about 15.0%. In an embodiment, theconcentration of silk in the solution ranges from about 0.5% to about10.0%. In an embodiment, the concentration of silk in the solutionranges from about 1.0% to about 5.0%. In an embodiment, an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure is applied directly to a fabric. Alternatively, silkmicrosphere and any additives may be used for coating a fabric. In anembodiment, additives can be added to an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure before coating(e.g., alcohols) to further enhance material properties. In anembodiment, a silk coating of the present disclosure can have a patternto optimize properties of the silk on the fabric. In an embodiment, acoating is applied to a fabric under tension and/or lax to varypenetration in to the fabric.

In an embodiment, a silk coating of the present disclosure can beapplied at the yarn level, followed by creation of a fabric once theyarn is coated. In an embodiment, an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure can be spuninto fibers to make a silk fabric and/or silk fabric blend with othermaterials known in the apparel industry.

In an embodiment, a method for silk coating a fabric includes immersionof the fabric in any of the aqueous solutions of pure silk fibroin-basedprotein fragments of the present disclosure. In an embodiment, a methodfor silk coating a fabric includes spraying. In an embodiment, a methodfor silk coating a fabric includes chemical vapor deposition. In anembodiment, a method for silk coating a fabric includes electrochemicalcoating. In an embodiment, a method for silk coating a fabric includesknife coating to spread any of the aqueous solutions of pure silkfibroin-based protein fragments of the present disclosure onto thefabric. The coated fabric may then be air dried, dried under heat/airflow, or cross-linked to the fabric surface. In an embodiment, a dryingprocess includes curing with additives and/or ambient condition.

According to aspects illustrated herein, methods for preparing aqueoussolutions of pure silk fibroin-based protein fragments are disclosed. Inan embodiment, at least one pure silk fibroin-based protein fragment(SPF) mixture solution having a specific average weight averagemolecular weight (MW) range and polydispersity is created. In anembodiment, at least SPF mixture solution having a MW range betweenabout 6 kDa and 16 kDa and a polydispersity range between about 1.5 andabout 3.0 is created. In an embodiment, at least one SPF mixturesolution having a MW between about 17 kDa and 38 kDa and apolydispersity range between about 1.5 and about 3.0 is created. In anembodiment, at least one SPF mixture solution having a MW range betweenabout 39 kDa and 80 kDa and a polydispersity range between about 1.5 andabout 3.0 is created.

According to aspects illustrated herein, there is disclosed acomposition that includes pure silk fibroin-based protein fragments thatare substantially devoid of sericin, wherein the composition has anaverage weight average molecular weight ranging from about 6 kDa toabout 16 kDa, wherein the composition has a polydispersity of betweenabout 1.5 and about 3.0, wherein the composition is substantiallyhomogenous, wherein the composition includes between 0 ppm and about 500ppm of inorganic residuals, and wherein the composition includes between0 ppm and about 500 ppm of organic residuals. In an embodiment, the puresilk fibroin-based protein fragments have between about 10 ppm and about300 ppm of lithium bromide residuals and between about 10 ppm and about100 ppm of sodium carbonate residuals. In an embodiment, the lithiumbromide residuals are measurable using a high-performance liquidchromatography lithium bromide assay, and the sodium carbonate residualsare measurable using a high-performance liquid chromatography sodiumcarbonate assay. In an embodiment, the composition further includes lessthan 10% water. In an embodiment, the composition is in the form of asolution. In an embodiment, the composition includes from about 0.1 wt %to about 30.0 wt % pure silk fibroin-based protein fragments. The puresilk fibroin-based protein fragments are stable in the solution for atleast 30 days. In an embodiment, the term “stable” refers to the absenceof spontaneous or gradual gelation, with no visible change in the coloror turbidity of the solution. In an embodiment, the term “stable” refersto no aggregation of fragments and therefore no increase in molecularweight over time. In an embodiment, the composition is in the form of anaqueous solution. In an embodiment, the composition is in the form of anorganic solution. The composition may be provided in a sealed container.In some embodiments, the composition further includes one or moremolecules selected from the group consisting of therapeutic agents,growth factors, antioxidants, proteins, vitamins, carbohydrates,polymers, nucleic acids, salts, acids, bases, biomolecules, glycosaminoglycans, polysaccharides, extracellular matrix molecules, metals, metalion, metal oxide, synthetic molecules, polyanhydrides, cells, fattyacids, fragrance, minerals, plants, plant extracts, preservatives andessential oils. In an embodiment, the added molecule or molecules arestable (i.e., retain activity over time) within the composition and canbe released at a desired rate. In an embodiment, the one or moremolecules is vitamin C or a derivative thereof. In an embodiment, thecomposition further includes an alpha hydroxy acid selected from thegroup consisting of glycolic acid, lactic acid, tartaric acid and citricacid. In an embodiment, the composition further includes hyaluronic acidor its salt form at a concentration of about 0.5% to about 10.0%. In anembodiment, the composition further includes at least one of zinc oxideor titanium dioxide. In an embodiment, the pure silk fibroin-basedprotein fragments in the composition are hypoallergenic. In anembodiment, the pure silk fibroin-based protein fragments arebiocompatible, non-sensitizing, and non-immunogenic.

According to aspects illustrated herein, there is disclosed acomposition that includes pure silk fibroin-based protein fragments thatare substantially devoid of sericin, wherein the composition has anaverage weight average molecular weight ranging from about 17 kDa toabout 38 kDa, wherein the composition has a polydispersity of betweenabout 1.5 and about 3.0, wherein the composition is substantiallyhomogenous, wherein the composition includes between 0 ppm and about 500ppm of inorganic residuals, and wherein the composition includes between0 ppm and about 500 ppm of organic residuals. In an embodiment, the puresilk fibroin-based protein fragments have between about 10 ppm and about300 ppm of lithium bromide residuals and between about 10 ppm and about100 ppm of sodium carbonate residuals. In an embodiment, the lithiumbromide residuals are measurable using a high-performance liquidchromatography lithium bromide assay, and the sodium carbonate residualsare measurable using a high-performance liquid chromatography sodiumcarbonate assay. In an embodiment, the composition further includes lessthan 10% water. In an embodiment, the composition is in the form of asolution. In an embodiment, the composition includes from about 0.1 wt %to about 30.0 wt % pure silk fibroin-based protein fragments. The puresilk fibroin-based protein fragments are stable in the solution for atleast 30 days. In an embodiment, the term “stable” refers to the absenceof spontaneous or gradual gelation, with no visible change in the coloror turbidity of the solution. In an embodiment, the term “stable” refersto no aggregation of fragments and therefore no increase in molecularweight over time. In an embodiment, the composition is in the form of anaqueous solution. In an embodiment, the composition is in the form of anorganic solution. The composition may be provided in a sealed container.In some embodiments, the composition further includes one or moremolecules selected from the group consisting of therapeutic agents,growth factors, antioxidants, proteins, vitamins, carbohydrates,polymers, nucleic acids, salts, acids, bases, biomolecules, glycosaminoglycans, polysaccharides, extracellular matrix molecules, metals, metalion, metal oxide, synthetic molecules, polyanhydrides, cells, fattyacids, fragrance, minerals, plants, plant extracts, preservatives andessential oils. In an embodiment, the added molecule or molecules arestable (i.e., retain activity over time) within the composition and canbe released at a desired rate. In an embodiment, the one or moremolecules is vitamin C or a derivative thereof. In an embodiment, thecomposition further includes an alpha hydroxy acid selected from thegroup consisting of glycolic acid, lactic acid, tartaric acid and citricacid. In an embodiment, the composition further includes hyaluronic acidor its salt form at a concentration of about 0.5% to about 10.0%. In anembodiment, the composition further includes at least one of zinc oxideor titanium dioxide. In an embodiment, the pure silk fibroin-basedprotein fragments in the composition are hypoallergenic. In anembodiment, the pure silk fibroin-based protein fragments arebiocompatible, non-sensitizing, and non-immunogenic.

According to aspects illustrated herein, there is disclosed acomposition that includes pure silk fibroin-based protein fragments thatare substantially devoid of sericin, wherein the composition has anaverage weight average molecular weight ranging from about 39 kDa toabout 80 kDa, wherein the composition has a polydispersity of betweenabout 1.5 and about 3.0, wherein the composition is substantiallyhomogenous, wherein the composition includes between 0 ppm and about 500ppm of inorganic residuals, and wherein the composition includes between0 ppm and about 500 ppm of organic residuals. In an embodiment, the puresilk fibroin-based protein fragments have between about 10 ppm and about300 ppm of lithium bromide residuals and between about 10 ppm and about100 ppm of sodium carbonate residuals. In an embodiment, the lithiumbromide residuals are measurable using a high-performance liquidchromatography lithium bromide assay, and the sodium carbonate residualsare measurable using a high-performance liquid chromatography sodiumcarbonate assay. In an embodiment, the composition further includes lessthan 10% water. In an embodiment, the composition is in the form of asolution. In an embodiment, the composition includes from about 0.1 wt %to about 30.0 wt % pure silk fibroin-based protein fragments. The puresilk fibroin-based protein fragments are stable in the solution for atleast 30 days. In an embodiment, the term “stable” refers to the absenceof spontaneous or gradual gelation, with no visible change in the coloror turbidity of the solution. In an embodiment, the term “stable” refersto no aggregation of fragments and therefore no increase in molecularweight over time. In an embodiment, the composition is in the form of anaqueous solution. In an embodiment, the composition is in the form of anorganic solution. The composition may be provided in a sealed container.In some embodiments, the composition further includes one or moremolecules selected from the group consisting of therapeutic agents,growth factors, antioxidants, proteins, vitamins, carbohydrates,polymers, nucleic acids, salts, acids, bases, biomolecules, glycosaminoglycans, polysaccharides, extracellular matrix molecules, metals, metalion, metal oxide, synthetic molecules, polyanhydrides, cells, fattyacids, fragrance, minerals, plants, plant extracts, preservatives andessential oils. In an embodiment, the added molecule or molecules arestable (i.e., retain activity over time) within the composition and canbe released at a desired rate. In an embodiment, the one or moremolecules is vitamin C or a derivative thereof. In an embodiment, thecomposition further includes an alpha hydroxy acid selected from thegroup consisting of glycolic acid, lactic acid, tartaric acid and citricacid. In an embodiment, the composition further includes hyaluronic acidor its salt form at a concentration of about 0.5% to about 10.0%. In anembodiment, the composition further includes at least one of zinc oxideor titanium dioxide. In an embodiment, the pure silk fibroin-basedprotein fragments in the composition are hypoallergenic. In anembodiment, the pure silk fibroin-based protein fragments arebiocompatible, non-sensitizing, and non-immunogenic.

According to aspects illustrated herein, there is disclosed a gel thatincludes pure silk fibroin-based protein fragments substantially devoidof sericin and comprising: an average weight average molecular weightranging from about 17 kDa to about 38 kDa; and a polydispersity ofbetween about 1.5 and about 3.0; and water from about 20 wt. % to about99.9 wt. %, wherein the gel includes between 0 ppm and 500 ppm ofinorganic residuals, and wherein the gel includes between 0 ppm and 500ppm of organic residuals. In an embodiment, the gel includes betweenabout 1.0% and about 50.0% crystalline protein domains. In anembodiment, the gel includes from about 0.1 wt. % to about 6.0 wt. % ofpure silk fibroin-based protein fragments. In an embodiment, the gel hasa pH from about 1.0 to about 7.0. In an embodiment, the gel furtherincludes from about 0.5 wt. % to about 20.0 wt. % of vitamin C or aderivative thereof. In an embodiment, the vitamin C or a derivativethereof remains stable within the gel for a period of from about 5 daysto about 5 years. In an embodiment, the vitamin C or a derivativethereof is stable within the gel so as to result in release of thevitamin C in a biologically active form. In an embodiment, the gelfurther includes an additive selected from the group consisting ofvitamin E, rosemary oil, rose oil, lemon juice, lemon grass oil andcaffeine. In an embodiment, the gel is packaged in an airtightcontainer. In an embodiment, the pure silk fibroin-based proteinfragments are hypoallergenic. In an embodiment, the gel has less than 10colony forming units per milliliter.

According to aspects illustrated herein, there is disclosed a method forpreparing an aqueous solution of pure silk fibroin-based proteinfragments having an average weight average molecular weight ranging fromabout 6 kDa to about 16 kDa, the method including the steps of:degumming a silk source by adding the silk source to a boiling (100° C.)aqueous solution of sodium carbonate for a treatment time of betweenabout 30 minutes to about 60 minutes; removing sericin from the solutionto produce a silk fibroin extract comprising non-detectable levels ofsericin; draining the solution from the silk fibroin extract; dissolvingthe silk fibroin extract in a solution of lithium bromide having astarting temperature upon placement of the silk fibroin extract in thelithium bromide solution that ranges from about 60° C. to about 140° C.;maintaining the solution of silk fibroin-lithium bromide in an ovenhaving a temperature of about 140° C. for a period of at least 1 hour;removing the lithium bromide from the silk fibroin extract; andproducing an aqueous solution of silk protein fragments, the aqueoussolution comprising: fragments having an average weight averagemolecular weight ranging from about 6 kDa to about 16 kDa, and whereinthe aqueous solution of pure silk fibroin-based protein fragmentscomprises a polydispersity of between about 1.5 and about 3.0. In anembodiment, the method includes the step of drying the silk fibroinextract prior to the dissolving step. In an embodiment, the amount oflithium bromide residuals in the aqueous solution can be measured usinga high-performance liquid chromatography lithium bromide assay. In anembodiment, the amount of sodium carbonate residuals in the aqueoussolution can be measured using a high-performance liquid chromatographysodium carbonate assay. In an embodiment, the method includes the stepof adding a therapeutic agent to the aqueous solution of pure silkfibroin-based protein fragments. In an embodiment, the method includesthe step of adding a molecule selected from one of an antioxidant or anenzyme to the aqueous solution of pure silk fibroin-based proteinfragments. In an embodiment, the method includes the step of adding avitamin to the aqueous solution of pure silk fibroin-based proteinfragments. In an embodiment, the vitamin is selected from one of vitaminC or a derivative thereof. In an embodiment, the method further includesthe step of adding an alpha hydroxy acid to the aqueous solution of puresilk fibroin-based protein fragments. In an embodiment, the alphahydroxy acid is selected from the group consisting of glycolic acid,lactic acid, tartaric acid and citric acid. In an embodiment, the methodfurther includes the step of adding hyaluronic acid at a concentrationof about 0.5% to about 10.0% to the aqueous solution of pure silkfibroin-based protein fragments. In an embodiment, the method furtherincludes the step of adding at least one of zinc oxide or titaniumdioxide to the aqueous solution of pure silk fibroin-based proteinfragments.

According to aspects illustrated herein, there is disclosed a method forpreparing an aqueous solution of pure silk fibroin-based proteinfragments having an average weight average molecular weight ranging fromabout 17 kDa to about 38 kDa, the method including the steps of: addinga silk source to a boiling (100° C.) aqueous solution of sodiumcarbonate for a treatment time of between about 30 minutes to about 60minutes so as to result in degumming; removing sericin from the solutionto produce a silk fibroin extract comprising non-detectable levels ofsericin; draining the solution from the silk fibroin extract; dissolvingthe silk fibroin extract in a solution of lithium bromide having astarting temperature upon placement of the silk fibroin extract in thelithium bromide solution that ranges from about 80° C. to about 140° C.;maintaining the solution of silk fibroin-lithium bromide in a dry ovenhaving a temperature in the range between about 60° C. to about 100° C.for a period of at least 1 hour; removing the lithium bromide from thesilk fibroin extract; and producing an aqueous solution of pure silkfibroin-based protein fragments, wherein the aqueous solution of puresilk fibroin-based protein fragments comprises lithium bromide residualsof between about 10 ppm and about 300 ppm, wherein the aqueous solutionof silk protein fragments comprises sodium carbonate residuals ofbetween about 10 ppm and about 100 ppm, wherein the aqueous solution ofpure silk fibroin-based protein fragments comprises fragments having anaverage weight average molecular weight ranging from about 17 kDa toabout 38 kDa, and wherein the aqueous solution of pure silkfibroin-based protein fragments comprises a polydispersity of betweenabout 1.5 and about 3.0. In an embodiment, the method includes the stepof drying the silk fibroin extract prior to the dissolving step. In anembodiment, the amount of lithium bromide residuals in the aqueoussolution can be measured using a high-performance liquid chromatographylithium bromide assay. In an embodiment, the amount of sodium carbonateresiduals in the aqueous solution can be measured using ahigh-performance liquid chromatography sodium carbonate assay. In anembodiment, the method includes the step of adding a therapeutic agentto the aqueous solution of pure silk fibroin-based protein fragments. Inan embodiment, the method includes the step of adding a moleculeselected from one of an antioxidant or an enzyme to the aqueous solutionof pure silk fibroin-based protein fragments. In an embodiment, themethod includes the step of adding a vitamin to the aqueous solution ofpure silk fibroin-based protein fragments. In an embodiment, the vitaminis selected from one of vitamin C or a derivative thereof. In anembodiment, the method further includes the step of adding an alphahydroxy acid to the aqueous solution of pure silk fibroin-based proteinfragments.

In an embodiment, the alpha hydroxy acid is selected from the groupconsisting of glycolic acid, lactic acid, tartaric acid and citric acid.In an embodiment, the method further includes the step of addinghyaluronic acid at a concentration of about 0.5% to about 10.0% to theaqueous solution of pure silk fibroin-based protein fragments. In anembodiment, the method further includes the step of adding at least oneof zinc oxide or titanium dioxide to the aqueous solution of pure silkfibroin-based protein fragments.

According to aspects illustrated herein, there is disclosed a method forpreparing an aqueous solution of pure silk fibroin-based proteinfragments having an average weight average molecular weight ranging fromabout 39 kDa to about 80 kDa, the method including the steps of: addinga silk source to a boiling (100° C.) aqueous solution of sodiumcarbonate for a treatment time of about 30 minutes so as to result indegumming; removing sericin from the solution to produce a silk fibroinextract comprising non-detectable levels of sericin; draining thesolution from the silk fibroin extract; dissolving the silk fibroinextract in a solution of lithium bromide having a starting temperatureupon placement of the silk fibroin extract in the lithium bromidesolution that ranges from about 80° C. to about 140° C.; maintaining thesolution of silk fibroin-lithium bromide in a dry oven having atemperature in the range between about 60° C. to about 100° C. for aperiod of at least 1 hour; removing the lithium bromide from the silkfibroin extract; and producing an aqueous solution of pure silkfibroin-based protein fragments, wherein the aqueous solution of puresilk fibroin-based protein fragments comprises lithium bromide residualsof between about 10 ppm and about 300 ppm, sodium carbonate residuals ofbetween about 10 ppm and about 100 ppm, fragments having an averageweight average molecular weight ranging from about 40 kDa to about 65kDa, and wherein the aqueous solution of pure silk fibroin-based proteinfragments comprises a polydispersity of between about 1.5 and about 3.0.In an embodiment, the method includes the step of drying the silkfibroin extract prior to the dissolving step. In an embodiment, theamount of lithium bromide residuals in the aqueous solution can bemeasured using a high-performance liquid chromatography lithium bromideassay. In an embodiment, the amount of sodium carbonate residuals in theaqueous solution can be measured using a high-performance liquidchromatography sodium carbonate assay. In an embodiment, the methodincludes the step of adding a therapeutic agent to the aqueous solutionof pure silk fibroin-based protein fragments. In an embodiment, themethod includes the step of adding a molecule selected from one of anantioxidant or an enzyme to the aqueous solution of pure silkfibroin-based protein fragments. In an embodiment, the method includesthe step of adding a vitamin to the aqueous solution of pure silkfibroin-based protein fragments. In an embodiment, the vitamin isselected from one of vitamin C or a derivative thereof. In anembodiment, the method further includes the step of adding an alphahydroxy acid to the aqueous solution of pure silk fibroin-based proteinfragments. In an embodiment, the alpha hydroxy acid is selected from thegroup consisting of glycolic acid, lactic acid, tartaric acid and citricacid. In an embodiment, the method further includes the step of addinghyaluronic acid at a concentration of about 0.5% to about 10.0% to theaqueous solution of pure silk fibroin-based protein fragments. In anembodiment, the method further includes the step of adding at least oneof zinc oxide or titanium dioxide to the aqueous solution of pure silkfibroin-based protein fragments.

According to aspects illustrated herein, a method is disclosed forproducing silk gels having entrapped molecules or therapeutic agentssuch as those listed in the following paragraphs. In an embodiment, atleast one molecule or therapeutic agent of interest is physicallyentrapped into a SPF mixture solution of the present disclosure duringprocessing into aqueous gels. An aqueous silk gel of the presentdisclosure can be used to release at least one molecule or therapeuticagent of interest.

According to aspects illustrated herein, pure silk fibroin-based proteinfragments from aqueous solutions of the present disclosure can be formedinto yarns and fabrics including for example, woven or weaved fabrics,and these fabrics can be used in textiles, as described above.

According to aspects illustrated herein, silk fabric manufactured fromSPF mixture solutions of the present disclosure are disclosed. In anembodiment, at least one molecule or therapeutic agent of interest isphysically entrapped into a SPF mixture solution of the presentdisclosure. A silk film of the present disclosure can be used to releaseat least one molecule or therapeutic agent of interest.

BRIEF DESCRIPTION OF THE DRAWINGS

The presently disclosed embodiments will be further explained withreference to the attached drawings. The drawings shown are notnecessarily to scale, with emphasis instead generally being placed uponillustrating the principles of the presently disclosed embodiments.

FIG. 1 is a flow chart showing various embodiments for producing puresilk fibroin-based protein fragments (SPFs) of the present disclosure.

FIG. 2 is a flow chart showing various parameters that can be modifiedduring the process of producing SPFs of the present disclosure duringthe extraction and the dissolution steps.

FIG. 3 is a photograph showing dry extracted silk fibroin.

FIG. 4 is a photograph showing an embodiment of a SPF in the form of asolution of the present disclosure.

FIGS. 5A-5D are photographs showing dissolved silk in room temperaturelithium bromide (LiBr) solutions dissolved in a 60° C. oven for 4 hours(sericin extraction temperature and time were varied).

FIGS. 6A-6D are photographs showing dissolved silk in room temperatureLiBr solutions dissolved in a 60° C. oven for 6 hours (sericinextraction temperature and time were varied).

FIGS. 7A-7D are photographs showing dissolved silk in room temperatureLiBr solutions dissolved in a 60° C. oven for 8 hours (sericinextraction temperature and time were varied).

FIGS. 8A-8D are photographs showing dissolved silk in room temperatureLiBr solutions dissolved in a 60° C. oven for 12 hours (sericinextraction temperature and time were varied).

FIGS. 9A-9D are photographs showing dissolved silk in room temperatureLiBr solutions dissolved in a 60° C. oven for 24 hours (sericinextraction temperature and time were varied).

FIGS. 10A-10D are photographs showing dissolved silk in room temperatureLiBr solutions dissolved in a 60° C. oven for 168/192 hours (sericinextraction temperature and time were varied).

FIGS. 11A-11C are photographs showing dissolved silk in room temperatureLiBr solutions dissolved in 60° C. oven for 1, 4, and 6 hours, wheresericin extraction was completed at 100° C. for 60 min.

FIGS. 12A-12D are photographs showing dissolved silk in 60° C. LiBrsolutions dissolved in a 60° C. oven for 1 hour (sericin extractiontemperature and time were varied).

FIGS. 13A-13D are photographs showing dissolved silk in 60° C. LiBrsolutions dissolved in a 60° C. oven for 4 hours (sericin extractiontemperature and time were varied).

FIGS. 14A-14D are photographs showing dissolved silk in 60° C. LiBrsolutions dissolved in a 60° C. oven for 6 hours (sericin extractiontemperature and time were varied).

FIGS. 15A-15D are photographs showing dissolved silk in 80° C. LiBrsolutions dissolved in a 60° C. oven for 1 hour (sericin extractiontemperature and time were varied).

FIGS. 16A-16D are photographs showing dissolved silk in 80° C. LiBrsolutions dissolved in a 60° C. oven for 4 hours (sericin extractiontemperature and time were varied).

FIGS. 17A-17D are photographs showing dissolved silk in 80° C. LiBrsolutions dissolved in a 60° C. oven for 4 hours (sericin extractiontemperature and time were varied).

FIGS. 18A-18D are photographs showing dissolved silk in 100° C. LiBrsolutions dissolved in a 60° C. oven for 1 hour (sericin extractiontemperature and time were varied).

FIGS. 19A-19D are photographs showing dissolved silk in 100° C. LiBrsolutions dissolved in a 60° C. oven for 4 hours (sericin extractiontemperature and time were varied).

FIGS. 20A-20D are photographs showing dissolved silk in 100° C. LiBrsolutions dissolved in a 60° C. oven for 6 hours (sericin extractiontemperature and time were varied).

FIGS. 21A-21D are photographs showing dissolved silk in 140° C. (boilingpoint for LiBr) LiBr solutions dissolved in a 60° C. oven for 1 hour(sericin extraction temperature and time were varied time).

FIGS. 22A-22D are photographs showing dissolved silk in 140° C. (boilingpoint for LiBr) LiBr solutions dissolved in a 60° C. oven for 4 hours(sericin extraction temperature and time were varied).

FIGS. 23A-23D are photographs showing dissolved silk in 140° C. (boilingpoint for LiBr) LiBr solutions dissolved in a 60° C. oven for 6 hours(sericin extraction temperature and time were varied).

FIGS. 24A-24D are photographs showing dissolved silk in 80° C. LiBrsolutions dissolved in a 80° C. oven for 1 hour (sericin extractiontemperature and time were varied).

FIGS. 25A-25D are photographs showing dissolved silk in 80° C. LiBrsolutions dissolved in a 80° C. oven for 4 hours (sericin extractiontemperature and time were varied).

FIGS. 26A-26D are photographs showing dissolved silk in 80° C. LiBrsolutions dissolved in a 80° C. oven for 6 hours (sericin extractiontemperature and time were varied).

FIGS. 27A-27D are photographs showing dissolved silk in 100° C. LiBrsolutions dissolved in a 100° C. oven for 1 hour (sericin extractiontemperature and time were varied).

FIGS. 28A-28D are photographs showing dissolved silk in 100° C. LiBrsolutions dissolved in a 100° C. oven for 4 hours (sericin extractiontemperature and time were varied).

FIGS. 29A-29D are photographs showing dissolved silk in 100° C. LiBrsolutions dissolved in a 100° C. oven for 6 hours (sericin extractiontemperature and time were varied).

FIGS. 30A-30D are photographs showing dissolved silk in 140° C. (boilingpoint for LiBr) LiBr solutions dissolved in a 120° C. oven for 1 hour(sericin extraction temperature and time were varied).

FIGS. 31A-31D are photographs showing dissolved silk in 140° C. (boilingpoint for LiBr) LiBr solutions dissolved in a 120° C. oven for 4 hours(sericin extraction temperature and time were varied).

FIG. 32A-32D are photographs showing dissolved silk in 140° C. (boilingpoint for LiBr) LiBr solutions dissolved in a 120° C. oven for 6 hours(sericin extraction temperature and time were varied).

FIG. 33 shows HPLC chromatograms from samples comprising vitamin C. FIG.33 shows peaks from (1) a chemically stabilized sample of vitamin C atambient conditions and (2) a sample of vitamin C taken after 1 hour atambient conditions without chemical stabilization to prevent oxidation,where degradation products are visible.

FIG. 34 is a table summarizing the LiBr and Sodium Carbonate (Na₂CO₃)concentration in silk protein solutions of the present disclosure.

FIG. 35 is a table summarizing the LiBr and Na₂CO₃ concentration in silkprotein solutions of the present disclosure.

FIG. 36 is a table summarizing the stability of vitamin C in chemicallystabilized solutions.

FIG. 37 is a table summarizing the Molecular Weights of silk proteinsolutions of the present disclosure.

FIGS. 38A and 38B are graphs representing the effect of extractionvolume on % mass loss.

FIG. 39 is a table summarizing the Molecular Weights of silk dissolvedfrom different concentrations of LiBr and from different extraction anddissolution sizes.

FIG. 40 is a graph summarizing the effect of Extraction Time onMolecular

Weight of silk processed under the conditions of 100° C. ExtractionTemperature, 100° C. LiBr and 100° C. Oven Dissolution (Oven/DissolutionTime was varied).

FIG. 41 is a graph summarizing the effect of Extraction Time onMolecular Weight of silk processed under the conditions of 100° C.Extraction Temperature, boiling LiBr and 60° C. Oven Dissolution(Oven/Dissolution Time was varied).

FIG. 42 is a graph summarizing the effect of Extraction Time onMolecular Weight of silk processed under the conditions of 100° C.Extraction Temperature, 60° C. LiBr and 60° C. Oven Dissolution(Oven/Dissolution Time was varied).

FIG. 43 is a graph summarizing the effect of Extraction Time onMolecular Weight of silk processed under the conditions of 100° C.Extraction Temperature, 80° C. LiBr and 80° C. Oven Dissolution(Oven/Dissolution Time was varied).

FIG. 44 is a graph summarizing the effect of Extraction Time onMolecular Weight of silk processed under the conditions of 100° C.Extraction Temperature, 80° C. LiBr and 60° C. Oven Dissolution(Oven/Dissolution Time was varied).

FIG. 45 is a graph summarizing the effect of Extraction Time onMolecular Weight of silk processed under the conditions of 100° C.Extraction Temperature, 100° C. LiBr and 60° C. Oven Dissolution(Oven/Dissolution Time was varied).

FIG. 46 is a graph summarizing the effect of Extraction Time onMolecular Weight of silk processed under the conditions of 100° C.Extraction Temperature, 140° C. LiBr and 140° C. Oven Dissolution(Oven/Dissolution Time was varied).

FIG. 47 is a graph summarizing the effect of Extraction Temperature onMolecular Weight of silk processed under the conditions of 60 minuteExtraction Time, 100° C. LiBr and 100° C. Oven Dissolution(Oven/Dissolution Time was varied).

FIG. 48 is a graph summarizing the effect of LiBr Temperature onMolecular Weight of silk processed under the conditions of 60 minuteExtraction Time, 100° C. Extraction Temperature and 60° C. OvenDissolution (Oven/Dissolution Time was varied).

FIG. 49 is a graph summarizing the effect of LiBr Temperature onMolecular Weight of silk processed under the conditions of 30 minuteExtraction Time, 100° C. Extraction Temperature and 60° C. OvenDissolution (Oven/Dissolution Time was varied).

FIG. 50 is a graph summarizing the effect of Oven/DissolutionTemperature on Molecular Weight of silk processed under the conditionsof 100° C. Extraction Temperature, 30 minute Extraction Time, and 100°C. Lithium Bromide (Oven/Dissolution Time was varied).

FIG. 51 is a graph summarizing the effect of Oven/DissolutionTemperature on Molecular Weight of silk processed under the conditionsof 100° C. Extraction Temperature, 60 minute Extraction Time, and 100°C. Lithium Bromide. (Oven/Dissolution Time was varied).

FIG. 52 is a graph summarizing the effect of Oven/DissolutionTemperature on Molecular Weight of silk processed under the conditionsof 100° C. Extraction Temperature, 60 minute Extraction Time, and 140°C. Lithium Bromide (Oven/Dissolution Time was varied).

FIG. 53 is a graph summarizing the effect of Oven/DissolutionTemperature on Molecular Weight of silk processed under the conditionsof 100° C. Extraction Temperature, 30 minute Extraction Time, and 140°C. Lithium Bromide (Oven/Dissolution Time was varied).

FIG. 54 is a graph summarizing the effect of Oven/DissolutionTemperature on Molecular Weight of silk processed under the conditionsof 100° C. Extraction Temperature, 60 minute Extraction Time, and 80° C.Lithium Bromide (Oven/Dissolution Time was varied).

FIG. 55 is a graph summarizing the Molecular Weights of silk processedunder varying conditions including Extraction Time, ExtractionTemperature, Lithium Bromide (LiBr) Temperature, Oven Temperature forDissolution, Oven Time for Dissolution.

FIG. 56 is a graph summarizing the Molecular Weights of silk processedunder conditions in which Oven/Dissolution Temperature is equal to LiBrTemperature.

FIG. 57A is a graph illustrating wetting time with spray coating.

FIG. 57B is a graph illustrating wetting time with stencil coating.

FIG. 57C is a graph illustrating wetting time with bath coating.

FIG. 57D is a graph illustrating wetting time with screen coating.

FIG. 58A is a graph illustrating absorption time with spray coating.

FIG. 58B is a graph illustrating absorption time with stencil coating.

FIG. 58C is a graph illustrating absorption time with bath coating.

FIG. 58D is a graph illustrating absorption time with screen coating.

FIG. 59A is a graph illustrating spreading speed with spray coating.

FIG. 59B is a graph illustrating spreading speed with stencil coating.

FIG. 59C is a graph illustrating spreading speed with bath coating.

FIG. 59D is a graph illustrating spreading speed with screen coating.

FIG. 60A is a graph illustrating accumulative one way transport indexwith spray coating.

FIG. 60B is a graph illustrating accumulative one way transport indexwith stencil coating.

FIG. 60C is a graph illustrating accumulative one way transport indexwith bath coating.

FIG. 60D is a graph illustrating accumulative one way transport indexwith screen coating.

FIG. 61A is a graph illustrating overall moisture management capabilitywith spray coating.

FIG. 61B is a graph illustrating overall moisture management capabilitywith stencil coating.

FIG. 61C is a graph illustrating overall moisture management capabilitywith bath coating.

FIG. 61D is a graph illustrating overall moisture management capabilitywith screen coating.

FIG. 62A is a graph illustrating wetting time top.

FIG. 62B is a graph illustrating wetting time bottom.

FIG. 63A is a graph illustrating top absorption rate.

FIG. 63B is a graph illustrating bottom absorption rate.

FIG. 64A is a graph illustrating top max wetted radius.

FIG. 64B is a graph illustrating bottom max wetted radius.

FIG. 65A is a graph illustrating top spreading speed.

FIG. 65B is a graph illustrating bottom spreading speed.

FIG. 66A is a graph illustrating accumulative one-way transport index.

FIG. 66B is a graph illustrating overall moisture management capability.

FIG. 67A is a graph illustrating wetting time of non-wicking finished.

FIG. 67B is a graph illustrating wetting time of semi-finished beforefinal setting.

FIG. 68A is a graph illustrating absorption time of non-wickingfinished.

FIG. 68B is a graph illustrating absorption time of semi-finished beforefinal setting.

FIG. 69A is a graph illustrating spreading speed of non-wickingfinished.

FIG. 69B is a graph illustrating spreading speed of semi-finished beforefinal setting.

FIG. 70A is a graph illustrating accumulative one way transport index ofnon-wicking finished.

FIG. 70B is a graph illustrating accumulative one way transport index ofsemi-finished before final setting.

FIG. 71A is a graph illustrating overall moisture management capabilityof non-wicking finished.

FIG. 71B is a graph illustrating overall moisture management capabilityof semi-finished before final setting.

FIG. 72A is a graph illustrating wetting time with spray coating.

FIG. 72B is a graph illustrating wetting time with stencil coating.

FIG. 72C is a graph illustrating wetting time with bath coating.

FIG. 73A is a graph illustrating absorption time with spray coating.

FIG. 73B is a graph illustrating absorption time with stencil coating.

FIG. 73C is a graph illustrating absorption time with bath coating.

FIG. 74A is a graph illustrating spreading speed with spray coating.

FIG. 74B is a graph illustrating spreading speed with stencil coating.

FIG. 74C is a graph illustrating spreading speed with bath coating.

FIG. 75A is a graph illustrating accumulative one way transport indexwith spray coating.

FIG. 75B is a graph illustrating accumulative one way transport indexwith stencil coating.

FIG. 75C is a graph illustrating accumulative one way transport indexwith bath coating.

FIG. 76A is a graph illustrating overall moisture management capabilitywith spray coating.

FIG. 76B is a graph illustrating overall moisture management capabilitywith stencil coating.

FIG. 76C is a graph illustrating overall moisture management capabilitywith bath coating.

FIG. 77A is a graph illustrating wetting time with 1% SFS.

FIG. 77B is a graph illustrating wetting time with 0.1% SFS.

FIG. 78A is a graph illustrating absorption time with 1% SFS.

FIG. 78B is a graph illustrating absorption time with 0.1% SFS.

FIG. 79A is a graph illustrating spreading speed with 1% SFS.

FIG. 79B is a graph illustrating spreading speed with 0.1% SFS.

FIG. 80A is a graph illustrating accumulative one way transport indexwith 1% SFS.

FIG. 80B is a graph illustrating accumulative one way transport indexwith 0.1% SFS.

FIG. 81A is a graph illustrating overall moisture management capabilitywith 1% SFS.

FIG. 81B is a graph illustrating overall moisture management capabilitywith 0.1% SFS.

FIG. 82A is a graph illustrating summary of wetting time top.

FIG. 82B is a graph illustrating summary of wetting time bottom.

FIG. 83A is a graph illustrating summary of top absorption rate.

FIG. 83B is a graph illustrating summary of bottom absorption rate.

FIG. 84A is a graph illustrating summary of top max wetted radius.

FIG. 84B is a graph illustrating summary of bottom wetted radius.

FIG. 85A is a graph illustrating summary of top spreading speed.

FIG. 85B is a graph illustrating summary of bottom spreading speed.

FIG. 86A is a graph illustrating summary of accumulative one-waytransport index.

FIG. 86B is a graph illustrating summary of overall moisture managementcapability.

FIG. 87 illustrates bacterial growth results.

FIG. 88 illustrates bacterial growth results.

FIG. 89 illustrates bacterial growth results.

FIG. 90 illustrates bacterial growth results.

FIG. 91 illustrates bacterial growth results.

FIG. 92 illustrates bacterial growth results.

FIG. 93 illustrates accumulative one-way transport index versus fabricwashing cycles.

FIG. 94 illustrates overall moisture management capability (OMMC) versusfabric washing cycles.

FIG. 95 illustrates wetting time at the top of the fabric versus fabricwashing cycles.

FIG. 96 illustrates wetting time at the bottom of the fabric versusfabric washing cycles.

FIG. 97 illustrates absorption rate at the top of the fabric versusfabric washing cycles.

FIG. 98 illustrates absorption rate at the bottom of the fabric versusfabric washing cycles.

FIG. 99 illustrates spreading speed at the top of the fabric versusfabric washing cycles.

FIG. 100 illustrates spreading speed at the bottom of the fabric versusfabric washing cycles.

FIG. 101 illustrates wetted radius at the top of the fabric versusfabric washing cycles.

FIG. 102 illustrates wetted radius at the bottom of the fabric versusfabric washing cycles.

FIG. 103 illustrates percent reduction in growth of Staphylococcusaureus ATCC 6538 versus fabric washing cycles.

FIG. 104 illustrates percent reduction in growth of Klebisiellapneumoniae ATCC 4354 versus fabric washing cycles.

FIG. 105 illustrates a scanning electron microscopy image of fabricsample FAB-01-BATH-B (first view).

FIG. 106 illustrates a scanning electron microscopy image of fabricsample FAB-01-BATH-B (second view).

FIG. 107 illustrates a scanning electron microscopy image of fabricsample FAB-01-BATH-B (third view).

FIG. 108 illustrates a scanning electron microscopy image of fabricsample FAB-01-BATH-B (fourth view).

FIG. 109 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-B (first view).

FIG. 110 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-B (second view).

FIG. 111 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-B (third view).

FIG. 112 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-B (fourth view).

FIG. 113 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-B (fifth view).

FIG. 114 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-B (sixth view).

FIG. 115 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-B (seventh view).

FIG. 116 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-C (first view).

FIG. 117 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-C (second view).

FIG. 118 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-C (third view).

FIG. 119 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-C (fourth view).

FIG. 120 illustrates a scanning electron microscopy image of fabricsample FAB-01-SPRAY-C (fifth view).

FIG. 121 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (first view).

FIG. 122 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (second view).

FIG. 123 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (third view).

FIG. 124 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (fourth view).

FIG. 125 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (fifth view).

FIG. 126 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (sixth view).

FIG. 127 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (seventh view).

FIG. 128 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (eighth view).

FIG. 129 illustrates a scanning electron microscopy image of fabricsample FAB-01-STEN-C (ninth view).

FIG. 130 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-B (first view).

FIG. 131 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-B (second view).

FIG. 132 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-B (third view).

FIG. 133 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-B (fourth view).

FIG. 134 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-B (fifth view).

FIG. 135 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-B (sixth view).

FIG. 136 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-B (seventh view).

FIG. 137 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (first view).

FIG. 138 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (second view).

FIG. 139 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (third view).

FIG. 140 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (fourth view).

FIG. 141 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (fifth view).

FIG. 142 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (sixth view).

FIG. 143 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (seventh view).

FIG. 144 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (eighth view).

FIG. 145 illustrates a scanning electron microscopy image of fabricsample FAB-10-BATH-C (ninth view).

FIG. 146 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (first view).

FIG. 147 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (second view).

FIG. 148 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (third view).

FIG. 149 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (fourth view).

FIG. 150 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (fifth view).

FIG. 151 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (sixth view).

FIG. 152 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (seventh view).

FIG. 153 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (eighth view).

FIG. 154 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-B (ninth view).

FIG. 155 illustrates a scanning electron microscopy image of fabricsample FAB-10-SPRAY-C.

FIG. 156 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (first view).

FIG. 157 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (second view).

FIG. 158 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (third view).

FIG. 159 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (fourth view).

FIG. 160 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (fifth view).

FIG. 161 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (sixth view).

FIG. 162 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (seventh view).

FIG. 163 illustrates a scanning electron microscopy image of fabricsample FAB-10-STEN-B (eighth view).

FIG. 164 illustrates a scanning electron microscopy image of a fabriccontrol sample (first view).

FIG. 165 illustrates a scanning electron microscopy image of a fabriccontrol sample (second view).

FIG. 166 illustrates a scanning electron microscopy image of a fabriccontrol sample (third view).

FIG. 167 illustrates a scanning electron microscopy image of a fabriccontrol sample (fourth view).

FIG. 168 illustrates a scanning electron microscopy image of film sampleFIL-01-BATH-B-01MYL (first view).

FIG. 169 illustrates a scanning electron microscopy image of film sampleFIL-01-BATH-B-01MYL (second view).

FIG. 170 illustrates a scanning electron microscopy image of film sampleFIL-01-BATH-B-01MYL (third view).

FIG. 171 illustrates a scanning electron microscopy image of film sampleFIL-01-BATH-B-01MYL (fourth view).

FIG. 172 illustrates a scanning electron microscopy image of film sampleFIL-01-BATH-B-01MYL (fifth view).

FIG. 173 illustrates a scanning electron microscopy image of film sampleFIL-01-BATH-B-01MYL (sixth view).

FIG. 174 illustrates a scanning electron microscopy image of film sampleFIL-01-BATH-B-01MYL (seventh view).

FIG. 175 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (first view).

FIG. 176 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (second view).

FIG. 177 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (third view).

FIG. 178 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (fourth view).

FIG. 179 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (fifth view).

FIG. 180 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (sixth view).

FIG. 181 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (seventh view).

FIG. 182 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-01MYL (eighth view).

FIG. 183 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-007MYL (first view).

FIG. 184 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-007MYL (second view).

FIG. 185 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-007MYL (third view).

FIG. 186 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-007MYL (fourth view).

FIG. 187 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-007MYL (fifth view).

FIG. 188 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-O1MYL_cross-section (first view).

FIG. 189 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-O1MYL_cross-section (second view).

FIG. 190 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-O1MYL_cross-section (third view).

FIG. 191 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-O1MYL_cross-section (fourth view).

FIG. 192 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-C-01MYL (first view).

FIG. 193 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-C-01MYL (second view).

FIG. 194 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-C-01MYL (third view).

FIG. 195 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-C-01MYL (fourth view).

FIG. 196 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-C-01MYL (fifth view).

FIG. 197 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-B-01-MYL (first view).

FIG. 198 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-B-01-MYL (second view).

FIG. 199 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-B-01-MYL (third view).

FIG. 200 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-B-01-MYL (fourth view).

FIG. 201 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-C-01-MYL (first view).

FIG. 202 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-C-01-MYL (second view).

FIG. 203 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-C-01-MYL (third view).

FIG. 204 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-C-01-MYL (fourth view).

FIG. 205 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-C-01-MYL (fifth view).

FIG. 206 illustrates a scanning electron microscopy image of film sampleFIL-01-STEN-C-01-MYL (sixth view).

FIG. 207 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-01MYL (first view).

FIG. 208 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-01MYL (second view).

FIG. 209 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-01MYL (third view).

FIG. 210 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-01MYL (fourth view).

FIG. 211 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-01MYL (fifth view).

FIG. 212 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-01MYL (sixth view).

FIG. 213 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-01MYL (seventh view).

FIG. 214 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-007MEL (first view).

FIG. 215 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-007MEL (second view).

FIG. 216 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-007MEL (third view).

FIG. 217 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-007MEL (fourth view).

FIG. 218 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-B-007MEL (fifth view).

FIG. 219 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-C-01MYL_cross-section (first view).

FIG. 220 illustrates a scanning electron microscopy image of film sampleFIL-10-SPRAY-B-01MYL (first view).

FIG. 221 illustrates a scanning electron microscopy image of film sampleFIL-10-SPRAY-B-01MYL (second view).

FIG. 222 illustrates a scanning electron microscopy image of film sampleFIL-10-SPRAY-B-01MYL (third view).

FIG. 223 illustrates a scanning electron microscopy image of film sampleFIL-10-SPRAY-B-01MYL (fourth view).

FIG. 224 illustrates a scanning electron microscopy image of film sampleFIL-10-SPRAY-B-01MYL (fifth view).

FIG. 225 illustrates a scanning electron microscopy image of film sampleFIL-10-SPRAY-B-01MYL (sixth view).

FIG. 226 illustrates a scanning electron microscopy image of film sampleFIL-BATH-C-01-MYL (first view).

FIG. 227 illustrates a scanning electron microscopy image of film sampleFIL-BATH-C-01-MYL (second view).

FIG. 228 illustrates a scanning electron microscopy image of film sampleFIL-BATH-C-01-MYL (third view).

FIG. 229 illustrates a scanning electron microscopy image of film sampleFIL-BATH-C-01-MYL (fourth view).

FIG. 230 illustrates a scanning electron microscopy image of film sampleFIL-BATH-C-01-MYL (fifth view).

FIG. 231 illustrates a scanning electron microscopy image of film sampleFIL-BATH-C-01-MYL (sixth view).

FIG. 232 illustrates a scanning electron microscopy image of film sampleMelinex Control (first view).

FIG. 233 illustrates a scanning electron microscopy image of film sampleMelinex Control (second view).

FIG. 234 illustrates a scanning electron microscopy image of film sampleMelinex Control (third view).

FIG. 235 illustrates a scanning electron microscopy image of film sampleMelinex Control (fourth view).

FIG. 236 illustrates a scanning electron microscopy image of film sampleMylar Control (first view).

FIG. 237 illustrates a scanning electron microscopy image of film sampleMylar Control (second view).

FIG. 238 illustrates a scanning electron microscopy image of film sampleMylar Control (third view).

FIG. 239 illustrates a scanning electron microscopy image of film sampleMylar Control (fourth view).

FIG. 240 illustrates a scanning electron microscopy image of film sampleMylar Control (fifth view).

FIG. 241 shows results from optical profiling measurements on the MylarControl sample taken at the top, location 1 (shiny side).

FIG. 242 shows results from optical profiling measurements on the MylarControl sample taken at the bottom, location 2 (more matte side).

FIG. 243 shows results from optical profiling measurements on theMelinex Control sample taken at the top, location 1.

FIG. 244 shows results from optical profiling measurements on theMelinex Control sample taken at the bottom, location 2.

FIG. 245 shows results from optical profiling measurements on sampleFIL-10-SPRAY-B-01MYL taken at the top, location 1.

FIG. 246 shows results from optical profiling measurements on sampleFIL-10-SPRAY-B-01MYL taken at the bottom, location 2.

FIG. 247 shows results from optical profiling measurements on sampleFIL-01-SPRAY-B-01MYL taken at the top, location 1.

FIG. 248 shows results from optical profiling measurements on sampleFIL-01-SPRAY-B-01MYL taken at the bottom, location 2.

FIG. 249 shows results from optical profiling measurements on sampleFIL-01-SPRAY-B-007MEL taken the top, location 1.

FIG. 250 shows results from optical profiling measurements on sampleFIL-01-SPRAY-B-007MEL taken at the bottom, location 2.

FIG. 251 shows results from optical profiling measurements on sampleFIL-01-SPRAY-C-01MYL taken at the top, location 1.

FIG. 252 shows results from optical profiling measurements on sampleFIL-01-SPRAY-C-01MYL taken at bottom, location 2

FIG. 253 shows results from optical profiling measurements on sampleFIL-01-STEN-B-01MYL taken at the top, location 1.

FIG. 254 shows results from optical profiling measurements on sampleFIL-01-STEN-B-01MYL taken at the bottom, location 2.

FIG. 255 shows results from optical profiling measurements on sampleFIL-01-STEN-C-01MYL taken at the top, location 1.

FIG. 256 shows results from optical profiling measurements on sampleFIL-01-STEN-C-01MYL taken at the bottom, location 2.

FIG. 257 shows results from optical profiling measurements on sampleFIL-10-BATH-B-01MYL taken at the top, location 1.

FIG. 258 shows results from optical profiling measurements on sampleFIL-10-BATH-B-01MYL taken at the bottom, Location 2.

FIG. 259 shows results from optical profiling measurements on sampleFIL-10-BATH-B-007MEL taken at the top, location 1.

FIG. 260 shows results from optical profiling measurements on sampleFIL-10-BATH-B-007MEL taken at the bottom, location 2.

FIG. 261 shows results from optical profiling measurements on sampleFIL-10-BATH-C-01MYL taken at top, location 1.

FIG. 262 shows results from optical profiling measurements on sampleFIL-10-BATH-C-01MYL taken at the bottom, location 2.

FIG. 263 shows results from optical profiling measurements on sampleFIL-01-BATH-B-01MYL taken at the top, location 1.

FIG. 264 shows results from optical profiling measurements on sampleFIL-01-BATH-B-01MYL taken at the bottom, location 2.

FIG. 265 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-O1MYL_cross-section.

FIG. 266 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-O1MYL_cross-section.

FIG. 267 illustrates a scanning electron microscopy image of film sampleFIL-01-SPRAY-B-O1MYL_cross-section.

FIG. 268 illustrates a scanning electron microscopy image of film sampleFIL-10-BATH-C-01MYL_cross-section.

FIG. 269 illustrates accumulative one-way transport index results fornatural fibers.

FIG. 270 illustrates overall moisture management capability for naturalfibers.

While the above-identified drawings set forth presently disclosedembodiments, other embodiments are also contemplated, as noted in thediscussion. This disclosure presents illustrative embodiments by way ofrepresentation and not limitation. Numerous other modifications andembodiments can be devised by those skilled in the art which fall withinthe scope and spirit of the principles of the presently disclosedembodiments.

DETAILED DESCRIPTION OF THE INVENTION

Provided herein are methods for producing pure and highly scalable silkprotein fragment (SPF) mixture solutions that may be used to coat atleast a portion of textiles or may be formed into usable fibers forweaving into yarn. The solutions are generated from raw pure intact silkprotein material and processed in order to remove any sericin andachieve the desired weight average molecular weight (MW) andpolydispersity of the fragment mixture. Select method parameters may bealtered to achieve distinct final silk protein fragment characteristicsdepending upon the intended use. The resulting final fragment solutionis pure silk protein fragments and water with PPM to non-detectablelevels of process contaminants. The concentration, size andpolydispersity of silk protein fragments in the solution may further bealtered depending upon the desired use and performance requirements. Inan embodiment, the pure silk fibroin-based protein fragments in thesolution are substantially devoid of sericin, have an average weightaverage molecular weight ranging from about 6 kDa to about 16 kDa, andhave a polydispersity ranging from about 1.5 and about 3.0. In anembodiment, the pure silk fibroin-based protein fragments in thesolution are substantially devoid of sericin, have an average weightaverage molecular weight ranging from about 17 kDa to about 38 kDa, andhave a polydispersity ranging from about 1.5 and about 3.0. In anembodiment, the pure silk fibroin-based protein fragments in thesolution are substantially devoid of sericin, have an average weightaverage molecular weight ranging from about 39 kDa to about 80 kDa, andhave a polydispersity ranging from about 1.5 and about 3.0. In anembodiment, the solutions may be used to generate articles, such as silkgels of varying gel and liquid consistencies by varying watercontent/concentration, or sold as a raw ingredient into the consumermarket.

As used herein, the terms “substantially sericin free” or “substantiallydevoid of sericin” refer to silk fibers in which a majority of thesericin protein has been removed. In an embodiment, silk fibroin that issubstantially devoid of sericin refers to silk fibroin having betweenabout 0.01% (w/w) and about 10.0% (w/w) sericin. In an embodiment, silkfibroin that is substantially devoid of sericin refers to silk fibroinhaving between about 0.01% (w/w) and about 9.0% (w/w) sericin. In anembodiment, silk fibroin that is substantially devoid of sericin refersto silk fibroin having between about 0.01% (w/w) and about 8.0% (w/w)sericin. In an embodiment, silk fibroin that is substantially devoid ofsericin refers to silk fibroin having between about 0.01% (w/w) andabout 7.0% (w/w) sericin. In an embodiment, silk fibroin that issubstantially devoid of sericin refers to silk fibroin having betweenabout 0.01% (w/w) and about 6.0% (w/w) sericin. In an embodiment, silkfibroin that is substantially devoid of sericin refers to silk fibroinhaving between about 0.01% (w/w) and about 5.0% (w/w) sericin. In anembodiment, silk fibroin that is substantially devoid of sericin refersto silk fibroin having between about 0% (w/w) and about 4.0% (w/w)sericin. In an embodiment, silk fibroin that is substantially devoid ofsericin refers to silk fibroin having between about 0.05% (w/w) andabout 4.0% (w/w) sericin. In an embodiment, silk fibroin that issubstantially devoid of sericin refers to silk fibroin having betweenabout 0.1% (w/w) and about 4.0% (w/w) sericin. In an embodiment, silkfibroin that is substantially devoid of sericin refers to silk fibroinhaving between about 0.5% (w/w) and about 4.0% (w/w) sericin. In anembodiment, silk fibroin that is substantially devoid of sericin refersto silk fibroin having between about 1.0% (w/w) and about 4.0% (w/w)sericin. In an embodiment, silk fibroin that is substantially devoid ofsericin refers to silk fibroin having between about 1.5% (w/w) and about4.0% (w/w) sericin. In an embodiment, silk fibroin that is substantiallydevoid of sericin refers to silk fibroin having between about 2.0% (w/w)and about 4.0% (w/w) sericin. In an embodiment, silk fibroin that issubstantially devoid of sericin refers to silk fibroin having betweenabout 2.5% (w/w) and about 4.0% (w/w) sericin. In an embodiment, silkfibroin that is substantially devoid of sericin refers to silk fibroinhaving a sericin content between about 0.01% (w/w) and about 0.1% (w/w).In an embodiment, silk fibroin that is substantially devoid of sericinrefers to silk fibroin having a sericin content below about 0.1% (w/w).In an embodiment, silk fibroin that is substantially devoid of sericinrefers to silk fibroin having a sericin content below about 0.05% (w/w).In an embodiment, when a silk source is added to a boiling (100° C.)aqueous solution of sodium carbonate for a treatment time of betweenabout 30 minutes to about 60 minutes, a degumming loss of about 26 wt. %to about 31 wt. % is obtained.

As used herein, the term “substantially homogeneous” may refer to puresilk fibroin-based protein fragments that are distributed in a normaldistribution about an identified molecular weight. As used herein, theterm “substantially homogeneous” may refer to an even distribution ofadditive, for example vitamin C, throughout a composition of the presentdisclosure.

As used herein, the term “substantially free of inorganic residuals”means that the composition exhibits residuals of 0.1% (w/w) or less. Inan embodiment, substantially free of inorganic residuals refers to acomposition that exhibits residuals of 0.05% (w/w) or less. In anembodiment, substantially free of inorganic residuals refers to acomposition that exhibits residuals of 0.01% (w/w) or less. In anembodiment, the amount of inorganic residuals is between 0 ppm(“non-detectable” or “ND”) and 1000 ppm. In an embodiment, the amount ofinorganic residuals is ND to about 500 ppm. In an embodiment, the amountof inorganic residuals is ND to about 400 ppm. In an embodiment, theamount of inorganic residuals is ND to about 300 ppm. In an embodiment,the amount of inorganic residuals is ND to about 200 ppm. In anembodiment, the amount of inorganic residuals is ND to about 100 ppm. Inan embodiment, the amount of inorganic residuals is between 10 ppm and1000 ppm.

As used herein, the term “substantially free of organic residuals” meansthat the composition exhibits residuals of 0.1% (w/w) or less. In anembodiment, substantially free of organic residuals refers to acomposition that exhibits residuals of 0.05% (w/w) or less. In anembodiment, substantially free of organic residuals refers to acomposition that exhibits residuals of 0.01% (w/w) or less. In anembodiment, the amount of organic residuals is between 0 ppm(“non-detectable” or “ND”) and 1000 ppm. In an embodiment, the amount oforganic residuals is ND to about 500 ppm. In an embodiment, the amountof organic residuals is ND to about 400 ppm. In an embodiment, theamount of organic residuals is ND to about 300 ppm. In an embodiment,the amount of organic residuals is ND to about 200 ppm. In anembodiment, the amount of organic residuals is ND to about 100 ppm. Inan embodiment, the amount of organic residuals is between 10 ppm and1000 ppm.

Compositions of the present disclosure exhibit “biocompatibility”meaning that the compositions are compatible with living tissue or aliving system by not being toxic, injurious, or physiologically reactiveand not causing immunological rejection. Such biocompatibility can beevidenced by participants topically applying compositions of the presentdisclosure on their skin for an extended period of time. In anembodiment, the extended period of time is about 3 days. In anembodiment, the extended period of time is about 7 days. In anembodiment, the extended period of time is about 14 days. In anembodiment, the extended period of time is about 21 days. In anembodiment, the extended period of time is about 30 days. In anembodiment, the extended period of time is selected from the groupconsisting of about 1 month, about 2 months, about 3 months, about 4months, about 5 months, about 6 months, about 7 months, about 8 months,about 9 months, about 10 months, about 11 months, about 12 months, andindefinitely.

Compositions of the present disclosure are “hypoallergenic” meaning thatthey are relatively unlikely to cause an allergic reaction. Suchhypoallergenicity can be evidenced by participants topically applyingcompositions of the present disclosure on their skin for an extendedperiod of time. In an embodiment, the extended period of time is about 3days. In an embodiment, the extended period of time is about 7 days. Inan embodiment, the extended period of time is about 14 days. In anembodiment, the extended period of time is about 21 days. In anembodiment, the extended period of time is about 30 days. In anembodiment, the extended period of time is selected from the groupconsisting of about 1 month, about 2 months, about 3 months, about 4months, about 5 months, about 6 months, about 7 months, about 8 months,about 9 months, about 10 months, about 11 months, about 12 months, andindefinitely.

As used herein, the term “washable” and “exhibiting washability” meansthat a silk coated fabric of the present disclosure is capable of beingwashed without shrinking, fading, or the like.

As used herein, the term “textile” refers to a flexible woven materialconsisting of a network of natural or artificial fibers often referredto as thread or yarn. In an embodiment, textiles can be used tofabricate clothing, shoes and bags. In an embodiment, textiles can beused to fabricate carpeting, upholstered furnishings, window shades,towels, and coverings for tables, beds, and other flat surfaces. In anembodiment, textiles can be used to fabricate flags, backpacks, tents,nets, handkerchiefs, balloons, kites, sails, and parachutes.

As used herein, the term “hand” refers to the feel of a fabric, whichmay be further described as the feeling of softness, crispness, dryness,silkiness, and combinations thereof. Fabric hand is also referred to as“drape.” A fabric with a hard hand is coarse, rough, and generally lesscomfortable for the wearer. A fabric with a soft hand is fluid andsmooth, such as fine silk or wool, and generally more comfortable forthe wearer. Fabric hand can be determined by comparison to collectionsof fabric samples, or by use of methods such as the Kawabata EvaluationSystem (KES) or the Fabric Assurance by Simple Testing (FAST) methods.Behera and Hari, Ind. J. Fibre & Textile Res., 1994, 19, 168-71.

As used herein, the term “yarn” refers to a single or multi-fiberconstruct.

As used herein, the term “bath coating” encompasses coating a fabric ina batch, immersing a fabric in a bath, and submerging a fabric in abath.

In an embodiment, the silk coating is applied using a bath process, ascreen (or stencil) process, a spray process, a silk-foam based process,and a roller based process.

In an embodiment, a fiber or a yarn comprises a synthetic fiber or yarn,including polyester, Mylar, cotton, nylon, polyester-polyurethanecopolymer, rayon, acetate, aramid (aromatic polyamide), acrylic, ingeo(polylactide), lurex (polyamide-polyester), olefin(polyethylene-polypropylene), and combinations thereof.

In an embodiment, a fiber or a yarn comprises a natural fiber or yarn,including alpaca fiber, alpaca fleece, alpaca wool, lama fiber, lamafleece, lama wool, cotton, cashmere and sheep fiber, sheep fleece, andsheep wool.

In an embodiment, a water-soluble silk coating may be used as anadhesive or binder for binding particles to fabrics or for bindingfabrics. In an embodiment, an article comprises a fabric bound toanother fabric using a silk coating. In an embodiment, an articlecomprises a fabric with particles bound to the fabric using a silkadhesive.

In an embodiment, the coating is applied to an article including afabric at the yarn level. In an embodiment, the coating is applied atthe fabric level. In an embodiment, the coating has a thickness selectedfrom the group consisting of about 5 nm, about 10 nm, about 15 nm, about20 nm, about 25 nm, about 50 nm, about 100 nm, about 200 nm, about 500nm, about 1 μm, about 5 μm, about 10 μm, and about 20 μm. In anembodiment, the coating has a thickness range selected from the groupconsisting of about 5 nm to about 100 nm, about 100 nm to about 200 nm,about 200 nm to about 500 nm, about 1 μm to about 2 μm, about 2 μm toabout 5 μm, about 5 μm to about 10 μm, and about 10 μm to about 20 μm.

In an embodiment, a fiber or a yarn is treated with a polymer, such aspolyglycolide (PGA), polyethylene glycols, copolymers of glycolide,glycolide/L-lactide copolymers (PGA/PLLA), glycolide/trimethylenecarbonate copolymers (PGA/TMC), polylactides (PLA), stereocopolymers ofPLA, poly-L-lactide (PLLA), poly-DL-lactide (PDLLA),L-lactide/DL-lactide copolymers, co-polymers of PLA,lactide/tetramethylglycolide copolymers, lactide/trimethylene carbonatecopolymers, lactide/δ-valerolactone copolymers, lactide/ε-caprolactonecopolymers, polydepsipeptides, PLA/polyethylene oxide copolymers,unsymmetrically 3,6-substituted poly-1,4-dioxane-2,5-diones,poly-β-hydroxybutyrate (PHBA), PHBA/β-hydroxyvalerate copolymers(PHBA/HVA), poly-β-hydroxypropionate (PHPA), poly-p-dioxanone (PDS),poly-δ-valerolactone, poly-ε-caprolactone, methylmethacrylate-N-vinylpyrrolidine copolymers, polyesteramides, polyesters of oxalic acid,polydihydropyrans, polyalkyl-2-cyanoacrylates, polyurethanes (PU),polyvinylalcohols (PVA), polypeptides, poly-β-malic acid (PMLA),poly-β-alkanoic acids, polyvinylalcohol (PVA), polyethyleneoxide (PEO),chitine polymers, polyethylene, polypropylene, polyasetal, polyamides,polyesters, polysulphone, polyether ether ketone, polyethyleneterephthalate, polycarbonate, polyaryl ether ketone, and polyetherketone ketone.

In an embodiment, the silk coating surface can be modified silk crystalsthat range in size from nm to μm.

The criterion for “visibility” is satisfied by any one of the following:a change in the surface character of the textile; the silk coating fillsthe interstices where the yarns intersect; or the silk coating blurs orobscures the weave.

In an embodiment, a silk based protein or fragment solution may beutilized to coat at least a portion of a fabric which can be used tocreate a textile. In an embodiment, a silk based protein or fragmentsolution may be weaved into yarn that can be used as a fabric in atextile. In an embodiment, a silk based protein or fragment solution maybe used to coat a fiber. In an embodiment, the invention provides anarticle comprising a silk based protein or fragment solution coating atleast a portion of a fabric or a textile. In an embodiment, theinvention provides an article comprising a silk based protein orfragment solution coating a yarn. In an embodiment, the inventionprovides an article comprising a silk based protein or fragment solutioncoating a fiber.

In an embodiment, a solution of the present disclosure is contacted withan additive, such as a therapeutic agent and/or a molecule. In anembodiment, molecules include, but are not limited to, antioxidants andenzymes. In an embodiment, molecules include, but are not limited to,ceramics, ceramic particles, metals, metal particles, polymer particles,inorganic particles, organic particles, selenium, ubiquinonederivatives, thiol-based antioxidants, saccharide-containingantioxidants, polyphenols, botanical extracts, caffeic acid, apigenin,pycnogenol, resveratrol, folic acid, vitamin B12, vitamin B6, vitaminB3, vitamin E, vitamin C and derivatives thereof, vitamin D, vitamin A,astaxathin, Lutein, lycopene, essential fatty acids (omegas 3 and 6),iron, zinc, magnesium, flavonoids (soy, Curcumin, Silymarin,Pycnongeol), growth factors, aloe, hyaluronic acid, extracellular matrixproteins, cells, nucleic acids, biomarkers, biological reagents, zincoxide, benzyol peroxide, retnoids, titanium, allergens in a known dose(for sensitization treatment), essential oils including, but not limitedto, lemongrass or rosemary oil, and fragrances. Therapeutic agentsinclude, but are not limited to, small molecules, drugs, proteins,peptides and nucleic acids. In an embodiment, a solution of the presentdisclosure is contacted with an allergen of known quantity prior toforming the article. Allergens include but are not limited to milk,eggs, peanuts, tree nuts, fish, shellfish, soy and wheat. Known doses ofallergen loaded within a silk article can be released at a known ratefor controlled exposure allergy study, tests and sensitizationtreatment.

In an embodiment, a solution of the present disclosure is used to createan article with microneedles by standard methods known to one in the artfor controlled delivery of molecules or therapeutic agents to or throughthe skin.

As used herein, the term “fibroin” includes silkworm fibroin and insector spider silk protein. In an embodiment, fibroin is obtained fromBombyx mori. In an embodiment, the spider silk protein is selected fromthe group consisting of swathing silk (Achniform gland silk), egg sacsilk (Cylindriform gland silk), egg case silk (Tubuliform silk),non-sticky dragline silk (Ampullate gland silk), attaching thread silk(Pyriform gland silk), sticky silk core fibers (Flagelliform glandsilk), and sticky silk outer fibers (Aggregate gland silk).

FIG. 1 is a flow chart showing various embodiments for producing puresilk fibroin-based protein fragments (SPFs) of the present disclosure.It should be understood that not all of the steps illustrated arenecessarily required to fabricate all silk solutions of the presentdisclosure. As illustrated in FIG. 1 , step A, cocoons (heat-treated ornon-heat-treated), silk fibers, silk powder or spider silk can be usedas the silk source. If starting from raw silk cocoons from Bombyx mori,the cocoons can be cut into small pieces, for example pieces ofapproximately equal size, step B1. The raw silk is then extracted andrinsed to remove any sericin, step C1a. This results in substantiallysericin free raw silk. In an embodiment, water is heated to atemperature between 84° C. and 100° C. (ideally boiling) and then Na₂CO₃(sodium carbonate) is added to the boiling water until the Na₂CO₃ iscompletely dissolved. The raw silk is added to the boiling water/Na₂CO₃(100° C.) and submerged for approximately 15-90 minutes, where boilingfor a longer time results in smaller silk protein fragments. In anembodiment, the water volume equals about 0.4×raw silk weight and theNa₂CO₃ volume equals about 0.848×raw silk weight. In an embodiment, thewater volume equals 0.1×raw silk weight and the Na₂CO₃ volume ismaintained at 2.12 g/L. This is demonstrated in FIG. 38A and FIG. 38Bsilk mass (x-axis) was varied in the same volume of extraction solution(i.e., the same volume of water and concentration of Na₂CO₃) achievingsericin removal (substantially sericin free) as demonstrated by anoverall silk mass loss of 26 to 31 percent (y-axis). Subsequently, thewater dissolved Na₂CO₃ solution is drained and excess water/Na₂CO₃ isremoved from the silk fibroin fibers (e.g., ring out the fibroin extractby hand, spin cycle using a machine, etc.). The resulting silk fibroinextract is rinsed with warm to hot water to remove any remainingadsorbed sericin or contaminate, typically at a temperature range ofabout 40° C. to about 80° C., changing the volume of water at least once(repeated for as many times as required). The resulting silk fibroinextract is a substantially sericin-depleted silk fibroin. In anembodiment, the resulting silk fibroin extract is rinsed with water at atemperature of about 60° C. In an embodiment, the volume of rinse waterfor each cycle equals 0.1 L to 0.2 L×raw silk weight. It may beadvantageous to agitate, turn or circulate the rinse water to maximizethe rinse effect. After rinsing, excess water is removed from theextracted silk fibroin fibers (e.g., ring out fibroin extract by hand orusing a machine). Alternatively, methods known to one skilled in the artsuch as pressure, temperature, or other reagents or combinations thereofmay be used for the purpose of sericin extraction. Alternatively, thesilk gland (100% sericin free silk protein) can be removed directly froma worm. This would result in liquid silk protein, without any alterationof the protein structure, free of sericin.

The extracted fibroin fibers are then allowed to dry completely. FIG. 3is a photograph showing dry extracted silk fibroin. Once dry, theextracted silk fibroin is dissolved using a solvent added to the silkfibroin at a temperature between ambient and boiling, step C1b. In anembodiment, the solvent is a solution of Lithium bromide (LiBr) (boilingfor LiBr is 140° C.). Alternatively, the extracted fibroin fibers arenot dried but wet and placed in the solvent; solvent concentration canthen be varied to achieve similar concentrations as to when adding driedsilk to the solvent. The final concentration of LiBr solvent can rangefrom 0.1M to 9.3M. FIG. 39 is a table summarizing the Molecular Weightsof silk dissolved from different concentrations of Lithium Bromide(LiBr) and from different extraction and dissolution sizes. Completedissolution of the extracted fibroin fibers can be achieved by varyingthe treatment time and temperature along with the concentration ofdissolving solvent. Other solvents may be used including, but notlimited to, phosphate phosphoric acid, calcium nitrate, calcium chloridesolution or other concentrated aqueous solutions of inorganic salts. Toensure complete dissolution, the silk fibers should be fully immersedwithin the already heated solvent solution and then maintained at atemperature ranging from about 60° C. to about 140° C. for 1-168 hrs. Inan embodiment, the silk fibers should be fully immersed within thesolvent solution and then placed into a dry oven at a temperature ofabout 100° C. for about 1 hour.

The temperature at which the silk fibroin extract is added to the LiBrsolution (or vice versa) has an effect on the time required tocompletely dissolve the fibroin and on the resulting molecular weightand polydispersity of the final SPF mixture solution. In an embodiment,silk solvent solution concentration is less than or equal to 20% w/v. Inaddition, agitation during introduction or dissolution may be used tofacilitate dissolution at varying temperatures and concentrations. Thetemperature of the LiBr solution will provide control over the silkprotein fragment mixture molecular weight and polydispersity created. Inan embodiment, a higher temperature will more quickly dissolve the silkoffering enhanced process scalability and mass production of silksolution. In an embodiment, using a LiBr solution heated to atemperature between 80° C.-140° C. reduces the time required in an ovenin order to achieve full dissolution. Varying time and temperature at orabove 60° C. of the dissolution solvent will alter and control the MWand polydispersity of the SPF mixture solutions formed from the originalmolecular weight of the native silk fibroin protein.

Alternatively, whole cocoons may be placed directly into a solvent, suchas LiBr, bypassing extraction, step B2. This requires subsequentfiltration of silk worm particles from the silk and solvent solution andsericin removal using methods know in the art for separating hydrophobicand hydrophilic proteins such as a column separation and/orchromatography, ion exchange, chemical precipitation with salt and/orpH, and or enzymatic digestion and filtration or extraction, all methodsare common examples and without limitation for standard proteinseparation methods, step C2. Non-heat treated cocoons with the silkwormremoved, may alternatively be placed into a solvent such as LiBr,bypassing extraction. The methods described above may be used forsericin separation, with the advantage that non-heat treated cocoonswill contain significantly less worm debris.

Dialysis may be used to remove the dissolution solvent from theresulting dissolved fibroin protein fragment solution by dialyzing thesolution against a volume of water, step E1. Pre-filtration prior todialysis is helpful to remove any debris (i.e., silk worm remnants) fromthe silk and LiBr solution, step D. In one example, a 3 μm or 5 μmfilter is used with a flow-rate of 200-300 mL/min to filter a 0.1% to1.0% silk-LiBr solution prior to dialysis and potential concentration ifdesired. A method disclosed herein, as described above, is to use timeand/or temperature to decrease the concentration from 9.3M LiBr to arange from 0.1M to 9.3M to facilitate filtration and downstreamdialysis, particularly when considering creating a scalable processmethod. Alternatively, without the use of additional time or temperate,a 9.3M LiBr-silk protein fragment solution may be diluted with water tofacilitate debris filtration and dialysis. The result of dissolution atthe desired time and temperate filtration is a translucent particle-freeroom temperature shelf-stable silk protein fragment-LiBr solution of aknown MW and polydispersity. It is advantageous to change the dialysiswater regularly until the solvent has been removed (e.g., change waterafter 1 hour, 4 hours, and then every 12 hours for a total of 6 waterchanges). The total number of water volume changes may be varied basedon the resulting concentration of solvent used for silk proteindissolution and fragmentation. After dialysis, the final silk solutionmaybe further filtered to remove any remaining debris (i.e., silk wormremnants).

Alternatively, Tangential Flow Filtration (TFF), which is a rapid andefficient method for the separation and purification of biomolecules,may be used to remove the solvent from the resulting dissolved fibroinsolution, step E2. TFF offers a highly pure aqueous silk proteinfragment solution and enables scalability of the process in order toproduce large volumes of the solution in a controlled and repeatablemanner. The silk and LiBr solution may be diluted prior to TFF (20% downto 0.1% silk in either water or LiBr). Pre-filtration as described aboveprior to TFF processing may maintain filter efficiency and potentiallyavoids the creation of silk gel boundary layers on the filter's surfaceas the result of the presence of debris particles. Pre-filtration priorto TFF is also helpful to remove any remaining debris (i.e., silk wormremnants) from the silk and LiBr solution that may cause spontaneous orlong-term gelation of the resulting water only solution, step D. TFF,recirculating or single pass, may be used for the creation of water-silkprotein fragment solutions ranging from 0.1% silk to 30.0% silk (morepreferably, 0.1%-6.0% silk). Different cutoff size TFF membranes may berequired based upon the desired concentration, molecular weight andpolydispersity of the silk protein fragment mixture in solution.Membranes ranging from 1-100 kDa may be necessary for varying molecularweight silk solutions created for example by varying the length ofextraction boil time or the time and temperate in dissolution solvent(e.g., LiBr). In an embodiment, a TFF 5 or 10 kDa membrane is used topurify the silk protein fragment mixture solution and to create thefinal desired silk-to-water ratio. As well, TFF single pass, TFF, andother methods known in the art, such as a falling film evaporator, maybe used to concentrate the solution following removal of the dissolutionsolvent (e.g., LiBr) (with resulting desired concentration ranging from0.1% to 30% silk). This can be used as an alternative to standard HFIPconcentration methods known in the art to create a water-based solution.A larger pore membrane could also be utilized to filter out small silkprotein fragments and to create a solution of higher molecular weightsilk with and/or without tighter polydispersity values. FIG. 37 is atable summarizing Molecular Weights for some embodiments of silk proteinsolutions of the present disclosure. Silk protein solution processingconditions were as follows: 100° C. extraction for 20 min, roomtemperature rinse, LiBr in 60° C. oven for 4-6 hours. FIGS. 40-49further demonstrate manipulation of extraction time, LiBr dissolutionconditions, and TFF processing and resultant example molecular weightsand polydispersities. These examples are not intended to be limiting,but rather to demonstrate the potential of specifying parameters forspecific molecular weight silk fragment solutions.

An assay for LiBr and Na₂CO₃ detection was performed using an HPLCsystem equipped with evaporative light scattering detector (ELSD). Thecalculation was performed by linear regression of the resulting peakareas for the analyte plotted against concentration. More than onesample of a number of formulations of the present disclosure was usedfor sample preparation and analysis. Generally, four samples ofdifferent formulations were weighed directly in a 10 mL volumetricflask.

The analytical method developed for the quantitation of Na₂CO₃ and LiBrin silk protein formulations was found to be linear in the range 10-165μg/mL, with RSD for injection precision as 2% and 1% for area and 0.38%and 0.19% for retention time for sodium carbonate and lithium bromiderespectively. The analytical method can be applied for the quantitativedetermination of sodium carbonate and lithium bromide in silk proteinformulations.

The final silk protein fragment solution, as shown in FIG. 4 , is puresilk protein fragments and water with PPM to undetectable levels ofparticulate debris and/or process contaminants, including LiBr andNa₂CO₃. FIG. 34 and FIG. 35 are tables summarizing LiBr and Na₂CO₃concentrations in solutions of the present disclosure. In FIG. 34 , theprocessing conditions included 100° C. extraction for 60 min, 60° C.rinse, 100° C. LiBr in 100° C. oven for 60 min. TFF conditions includingpressure differential and number of dia-filtration volumes were varied.In FIG. 35 , the processing conditions included 100° C. boil for 60 min,60° C. rinse, LiBr in 60° C. oven for 4-6 hours. In an embodiment, a SPFcomposition of the present disclosure is not soluble in an aqueoussolution due to the crystallinity of the protein. In an embodiment, aSPF composition of the present disclosure is soluble in an aqueoussolution. In an embodiment, the SPFs of a composition of the presentdisclosure include a crystalline portion of about two-thirds and anamorphous region of about one-third. In an embodiment, the SPFs of acomposition of the present disclosure include a crystalline portion ofabout one-half and an amorphous region of about one-half. In anembodiment, the SPFs of a composition of the present disclosure includea 99% crystalline portion and a 1% amorphous region. In an embodiment,the SPFs of a composition of the present disclosure include a 95%crystalline portion and a 5% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 90%crystalline portion and a 10% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 85%crystalline portion and a 15% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 80%crystalline portion and a 20% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 75%crystalline portion and a 25% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 70%crystalline portion and a 30% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 65%crystalline portion and a 35% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 60%crystalline portion and a 40% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 50%crystalline portion and a 50% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 40%crystalline portion and a 60% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 35%crystalline portion and a 65% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 30%crystalline portion and a 70% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 25%crystalline portion and a 75% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 20%crystalline portion and a 80% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 15%crystalline portion and a 85% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 10%crystalline portion and a 90% amorphous region. In an embodiment, theSPFs of a composition of the present disclosure include a 5% crystallineportion and a 90% amorphous region. In an embodiment, the SPFs of acomposition of the present disclosure include a 1% crystalline portionand a 99% amorphous region.

A unique feature of the SPF compositions of the present disclosure areshelf stability (they will not slowly or spontaneously gel when storedin an aqueous solution and there is no aggregation of fragments andtherefore no increase in molecular weight over time), from 10 days to 3years depending on storage conditions, percent silk, and number ofshipments and shipment conditions. Additionally pH may be altered toextend shelf-life and/or support shipping conditions by preventingpremature folding and aggregation of the silk. In an embodiment, a SPFsolution composition of the present disclosure has a shelf stability forup to 2 weeks at room temperature (RT). In an embodiment, a SPF solutioncomposition of the present disclosure has a shelf stability for up to 4weeks at RT. In an embodiment, a SPF solution composition of the presentdisclosure has a shelf stability for up to 6 weeks at RT. In anembodiment, a SPF solution composition of the present disclosure has ashelf stability for up to 8 weeks at RT. In an embodiment, a SPFsolution composition of the present disclosure has a shelf stability forup to 10 weeks at RT. In an embodiment, a SPF solution composition ofthe present disclosure has a shelf stability for up to 12 weeks at RT.In an embodiment, a SPF solution composition of the present disclosurehas a shelf stability ranging from about 4 weeks to about 52 weeks atRT. Table 1 below shows shelf stability test results for embodiments ofSPF compositions of the present disclosure.

TABLE 1 Shelf Stability of SPF Compositions of the Present Disclosure %Silk Temperature Time to Gelation 2 RT 4 weeks 2 4 C. >9 weeks  4 RT 4weeks 4 4 C. >9 weeks  6 RT 2 weeks 6 4 C. >9 weeks 

A silk fragment-water solution of the present disclosure can besterilized following standard methods in the art not limited tofiltration, heat, radiation or e-beam. It is anticipated that the silkprotein fragment mixture, because of its shorter protein polymer length,will withstand sterilization better than intact silk protein solutionsdescribed in the art. Additionally, silk articles created from the SPFmixtures described herein may be sterilized as appropriate toapplication.

FIG. 2 is a flow chart showing various parameters that can be modifiedduring the process of producing a silk protein fragment solution of thepresent disclosure during the extraction and the dissolution steps.Select method parameters may be altered to achieve distinct finalsolution characteristics depending upon the intended use, e.g.,molecular weight and polydispersity. It should be understood that notall of the steps illustrated are necessarily required to fabricate allsilk solutions of the present disclosure.

In an embodiment, a process for producing a silk protein fragmentsolution of the present disclosure includes forming pieces of silkcocoons from the Bombyx mori silk worm; extracting the pieces at about100° C. in a solution of water and Na₂CO₃ for about 60 minutes, whereina volume of the water equals about 0.4×raw silk weight and the amount ofNa₂CO₃ is about 0.848×the weight of the pieces to form a silk fibroinextract; triple rinsing the silk fibroin extract at about 60° C. forabout 20 minutes per rinse in a volume of rinse water, wherein the rinsewater for each cycle equals about 0.2 L×the weight of the pieces;removing excess water from the silk fibroin extract; drying the silkfibroin extract; dissolving the dry silk fibroin extract in a LiBrsolution, wherein the LiBr solution is first heated to about 100° C. tocreate a silk and LiBr solution and maintained; placing the silk andLiBr solution in a dry oven at about 100° C. for about 60 minutes toachieve complete dissolution and further fragmentation of the nativesilk protein structure into mixture with desired molecular weight andpolydispersity; filtering the solution to remove any remaining debrisfrom the silkworm; diluting the solution with water to result in a 1%silk solution; and removing solvent from the solution using TangentialFlow Filtration (TFF). In an embodiment, a 10 kDa membrane is utilizedto purify the silk solution and create the final desired silk-to-waterratio. TFF can then be used to further concentrate the pure silksolution to a concentration of 2% silk to water.

Each process step from raw cocoons to dialysis is scalable to increaseefficiency in manufacturing. Whole cocoons are currently purchased asthe raw material, but pre-cleaned cocoons or non-heat treated cocoons,where worm removal leaves minimal debris, have also been used. Cuttingand cleaning the cocoons is a manual process, however for scalabilitythis process could be made less labor intensive by, for example, usingan automated machine in combination with compressed air to remove theworm and any particulates, or using a cutting mill to cut the cocoonsinto smaller pieces. The extraction step, currently performed in smallbatches, could be completed in a larger vessel, for example anindustrial washing machine where temperatures at or in between 60° C. to100° C. can be maintained. The rinsing step could also be completed inthe industrial washing machine, eliminating the manual rinse cycles.Dissolution of the silk in LiBr solution could occur in a vessel otherthan a convection oven, for example a stirred tank reactor. Dialyzingthe silk through a series of water changes is a manual and timeintensive process, which could be accelerated by changing certainparameters, for example diluting the silk solution prior to dialysis.The dialysis process could be scaled for manufacturing by usingsemi-automated equipment, for example a tangential flow filtrationsystem.

Varying extraction (i.e., time and temperature), LiBr (i.e., temperatureof LiBr solution when added to silk fibroin extract or vice versa) anddissolution (i.e., time and temperature) parameters results in solventand silk solutions with different viscosities, homogeneities, and colors(see FIGS. 5-32 ). Increasing the temperature for extraction,lengthening the extraction time, using a higher temperature LiBrsolution at emersion and over time when dissolving the silk andincreasing the time at temperature (e.g., in an oven as shown here, oran alternative heat source) all resulted in less viscous and morehomogeneous solvent and silk solutions. While almost all parametersresulted in a viable silk solution, methods that allow completedissolution to be achieved in fewer than 4 to 6 hours are preferred forprocess scalability.

FIGS. 5-10 show photographs of four different silk extractioncombinations tested: 90° C. 30 min, 90° C. 60 min, 100° C. 30 min, and100° C. 60 min. Briefly, 9.3 M LiBr was prepared and allowed to sit atroom temperature for at least 30 minutes. 5 mL of LiBr solution wasadded to 1.25 g of silk and placed in the 60° C. oven. Samples from eachset were removed at 4, 6, 8, 12, 24, 168 and 192 hours. The remainingsample was photographed.

FIGS. 11-23 show photographs of four different silk extractioncombinations tested: 90° C. 30 min, 90° C. 60 min, 100° C. 30 min, and100° C. 60 min. Briefly, 9.3 M LiBr solution was heated to one of fourtemperatures: 60° C., 80° C., 100° C. or boiling. 5 mL of hot LiBrsolution was added to 1.25 g of silk and placed in the 60° C. oven.Samples from each set were removed at 1, 4 and 6 hours. The remainingsample was photographed.

FIGS. 24-32 show photographs of four different silk extractioncombinations tested: Four different silk extraction combinations wereused: 90° C. 30 min, 90° C. 60 min, 100° C. 30 min, and 100° C. 60 min.Briefly, 9.3 M LiBr solution was heated to one of four temperatures: 60°C., 80° C., 100° C. or boiling. 5 mL of hot LiBr solution was added to1.25 g of silk and placed in the oven at the same temperature of theLiBr. Samples from each set were removed at 1, 4 and 6 hours. 1 mL ofeach sample was added to 7.5 mL of 9.3 M LiBr and refrigerated forviscosity testing. The remaining sample was photographed.

Molecular weight of the silk protein fragments may be controlled basedupon the specific parameters utilized during the extraction step,including extraction time and temperature; specific parameters utilizedduring the dissolution step, including the LiBr temperature at the timeof submersion of the silk in to the lithium bromide and time that thesolution is maintained at specific temperatures; and specific parametersutilized during the filtration step. By controlling process parametersusing the disclosed methods, it is possible to create SPF mixturesolutions with polydispersity equal to or lower than 2.5 at a variety ofdifferent molecular weight ranging from 5 kDa to 200 kDa, morepreferably between 10 kDa and 80 kDA. By altering process parameters toachieve silk solutions with different molecular weights, a range offragment mixture end products, with desired polydispersity of equal toor less than 2.5 may be targeted based upon the desired performancerequirements. Additionally, SPF mixture solutions with a polydispersityof greater than 2.5 can be achieved. Further, two solutions withdifferent average molecular weights and polydispersities can be mixed tocreate combination solutions. Alternatively, a liquid silk gland (100%sericin free silk protein) that has been removed directly from a wormcould be used in combination with any of the SPF mixture solutions ofthe present disclosure. Molecular weight of the pure silk fibroin-basedprotein fragment composition was determined using High Pressure LiquidChromatography (HPLC) with a Refractive Index Detector (RID).Polydispersity was calculated using Cirrus GPC Online GPC/SEC SoftwareVersion 3.3 (Agilent).

Parameters were varied during the processing of raw silk cocoons intosilk solution. Varying these parameters affected the MW of the resultingsilk solution. Parameters manipulated included (i) time and temperatureof extraction, (ii) temperature of LiBr, (iii) temperature ofdissolution oven, and (iv) dissolution time. Molecular weight wasdetermined with mass spec as shown in FIGS. 40-54 .

Experiments were carried out to determine the effect of varying theextraction time. FIGS. 40-46 are graphs showing these results, andTables 2-8 summarize the results. Below is a summary:

-   -   A sericin extraction time of 30 minutes resulted in larger MW        than a sericin extraction time of 60 minutes    -   MW decreases with time in the oven    -   140° C. LiBr and oven resulted in the low end of the confidence        interval to be below a MW of 9500 Da    -   30 min extraction at the 1 hour and 4 hour time points have        undigested silk    -   30 min extraction at the 1 hour time point resulted in a        significantly high molecular weight with the low end of the        confidence interval being 35,000 Da    -   The range of MW reached for the high end of the confidence        interval was 18000 to 216000 Da (important for offering        solutions with specified upper limit)

TABLE 2 The effect of extraction time (30 min vs 60 min) on molecularweight of silk processed under the conditions of 100° C. ExtractionTemperature, 100° C. Lithium Bromide (LiBr) and 100° C. Oven Dissolution(Oven/Dissolution Time was varied). Boil Time Oven Time Average Mw Stddev Confidence Interval PD 30 1 57247 12780 35093 93387 1.63 60 1 315201387 11633 85407 2.71 30 4 40973 2632 14268 117658 2.87 60 4 25082 124810520 59803 2.38 30 6 25604 1405 10252 63943 2.50 60 6 20980 1262 1007343695 2.08

TABLE 3 The effect of extraction time (30 min vs 60 min) on molecularweight of silk processed under the conditions of 100° C. ExtractionTemperature, boiling Lithium Bromide (LiBr) and 60° C. Oven Dissolutionfor 4 hr. Std Sample Boil Time Average Mw dev Confidence Interval PD 30min, 4 hr 30 49656 4580 17306 142478 2.87 60 min, 4 hr 60 30042 153611183 80705 2.69

TABLE 4 The effect of extraction time (30 min vs 60 min) on molecularweight of silk processed under the conditions of 100° C. ExtractionTemperature, 60° C. Lithium Bromide (LiBr) and 60° C. Oven Dissolution(Oven/Dissolution Time was varied). Oven Average Std Confidence SampleBoil Time Time Mw dev Interval PD 30 min, 1 hr 30 1 58436 22201 1538092.63 60 min, 1 hr 60 1 31700 11931 84224 2.66 30 min, 4 hr 30 4 61956.513337 21463 178847 2.89 60 min, 4 hr 60 4 25578.5 2446 9979 65564 2.56

TABLE 5 The effect of extraction time (30 min vs 60 min) on molecularweight of silk processed under the conditions of 100° C. ExtractionTemperature, 80° C. Lithium Bromide (LiBr) and 80° C. Oven Dissolutionfor 6 hr. Average Std Sample Boil Time Mw dev Confidence Interval PD 30min, 6 hr 30 63510 18693 215775 3.40 60 min, 6 hr 60 25164 238 963765706 2.61

TABLE 6 The effect of extraction time (30 min vs 60 min) on molecularweight of silk processed under the conditions of 100° C. ExtractionTemperature, 80° C. Lithium Bromide (LiBr) and 60° C. Oven Dissolution(Oven/Dissolution Time was varied). Oven Average Confidence Sample BoilTime Time Mw Std dev Interval PD 30 min, 4 hr 30 4 59202 14028 19073183760 3.10 60 min, 4 hr 60 4 26312.5 637 10266 67442 2.56 30 min, 6 hr30 6 46824 18076 121293 2.59 60 min, 6 hr 60 6 26353 10168 68302 2.59

TABLE 7 The effect of extraction time (30 min vs 60 min) on molecularweight of silk processed under the conditions of 100° C. ExtractionTemperature, 100° C. Lithium Bromide (LiBr) and 60° C. Oven Dissolution(Oven/Dissolution Time was varied). Boil Oven Average Std ConfidenceSample Time Time Mw dev Interval PD 30 min, 4 hr 30 4 47853 19758 1159002.42 60 min, 4 hr 60 4 25082 1248 10520  59804 2.38 30 min, 6 hr 30 655421 8992 19153 160366 2.89 60 min, 6 hr 60 6 20980 1262 10073  436942.08

TABLE 8 The effect of extraction time (30 min vs 60 min) on molecularweight of silk processed under the conditions of 100° C. ExtractionTemperature, 140° C. Lithium Bromide (LiBr) and 140° C. Oven Dissolution(Oven/Dissolution Time was varied). Boil Oven Average Std ConfidenceSample Time Time Mw dev Interval PD 30 min, 4 hr 30 4 9024.5 1102 449318127 2.00865 60 min, 4 hr 60 4 15548 6954 34762 2.2358  30 min, 6 hr 306 13021 5987 28319 2.1749  60 min, 6 hr 60 6 10888 5364 22100 2.0298 

Experiments were carried out to determine the effect of varying theextraction temperature. FIG. 47 is a graph showing these results, andTable 9 summarizes the results. Below is a summary:

-   -   Sericin extraction at 90° C. resulted in higher MW than sericin        extraction at 100° C. extraction    -   Both 90° C. and 100° C. show decreasing MW over time in the oven

TABLE 9 The effect of extraction temperature (90° C. vs. 100° C.) onmolecular weight of silk processed under the conditions of 60 min.Extraction Temperature, 100° C. Lithium Bromide (LiBr) and 100° C. OvenDissolution (Oven/Dissolution Time was varied). Boil Oven Average StdConfidence Sample Time Time Mw dev Interval PD  90 C., 4 hr 60 4 373084204 13368 104119 2.79 100 C., 4 hr 60 4 25082 1248 10520  59804 2.38 90 C., 6 hr 60 6 34224 1135 12717  92100 2.69 100 C., 6 hr 60 6 209801262 10073  43694 2.08

Experiments were carried out to determine the effect of varying theLithium Bromide (LiBr) temperature when added to silk. FIGS. 48-49 aregraphs showing these results, and Tables 10-11 summarize the results.Below is a summary:

-   -   No impact on MW or confidence interval (all CI˜10500-6500 Da)    -   Studies illustrated that the temperature of LiBr-silk        dissolution, as LiBr is added and begins dissolving, rapidly        drops below the original LiBr temperature due to the majority of        the mass being silk at room temp

TABLE 10 The effect of Lithium Bromide (LiBr) temperature on molecularweight of silk processed under the conditions of 60 min. ExtractionTime., 100° C. Extraction Temperature and 60° C. Oven Dissolution(Oven/Dissolution Time was varied). LiBr Temp Oven Average StdConfidence Sample (° C.) Time Mw dev Interval PD  60 C. LiBr, 1 hr  60 131700 11931 84223 2.66 100 C. LiBr, 1 hr 100 1 27907  200 10735 725521.60 RT LiBr, 4 hr RT 4 29217 1082 10789 79119 2.71  60 C. LiBr, 4 hr 60 4 25578 2445  9978 65564 2.56  80 C. LiBr, 4 hr  80 4 26312  63710265 67441 2.56 100 C. LiBr, 4 hr 100 4 27681 1729 11279 67931 2.45Boil LiBr, 4 hr Boil 4 30042 1535 11183 80704 2.69 RT LiBr, 6 hr RT 626543 1893 10783 65332 2.46  80 C. LiBr, 6 hr  80 6 26353 10167 683012.59 100 C. LiBr, 6 hr 100 6 27150  916 11020 66889 2.46

TABLE 11 The effect of Lithium Bromide (LiBr) temperature on molecularweight of silk processed under the conditions of 30 min. ExtractionTime, 100° C. Extraction Temperature and 60° C. Oven Dissolution(Oven/Dissolution Time was varied). LiBr Temp Oven Average StdConfidence Sample (° C.) Time Mw dev Interval PD  60 C. LiBr, 4 hr  60 461956 13336 21463 178847 2.89  80 C. LiBr, 4 hr  80 4 59202 14027 19073183760 3.10 100 C. LiBr, 4 hr 100 4 47853 19757 115899 2.42  80 C. LiBr,6 hr  80 6 46824 18075 121292 2.59 100 C. LiBr, 6 hr 100 6 55421  899119152 160366 2.89

Experiments were carried out to determine the effect of varying theoven/dissolution temperature. FIGS. 50-54 are graphs showing theseresults, and Tables 12-16 summarize the results. Below is a summary:

-   -   Oven temperature has less of an effect on 60 min extracted silk        than 30 min extracted silk. Without wishing to be bound by        theory, it is believed that the 30 min silk is less degraded        during extraction and therefore the oven temperature has more of        an effect on the larger MW, less degraded portion of the silk.    -   For 60° C. vs. 140° C. oven the 30 min extracted silk showed a        very significant effect of lower MW at higher oven temp, while        60 min extracted silk had an effect but much less    -   The 140° C. oven resulted in a low end in the confidence        interval at 6000 Da

TABLE 12 The effect of oven/dissolution temperature on molecular weightof silk processed under the conditions of 100° C. ExtractionTemperature, 30 min. Extraction Time, and 100° C. Lithium Bromide (LiBr)(Oven/Dissolution Time was varied). Oven Boil Temp Oven Average StdConfidence Time (° C.) Time Mw dev Interval PD 30  60 4 47853 19758115900 2.42 30 100 4 40973 2632 14268 117658 2.87 30  60 6 55421 899219153 160366 2.89 30 100 6 25604 1405 10252  63943 2.50

TABLE 13 The effect of oven/dissolution temperature on molecular weightof silk processed under the conditions of 100° C. ExtractionTemperature, 60 min. Extraction Time, and 100° C. Lithium Bromide (LiBr)(Oven/Dissolution Time was varied). Oven Boil Temp Oven Average StdConfidence Time (° C.) Time Mw dev Interval PD 60  60 1 27908  200 1073572552 2.60 60 100 1 31520 1387 11633 85407 2.71 60  60 4 27681 173011279 72552 2.62 60 100 4 25082 1248 10520 59803 2.38 60  60 6 27150 916 11020 66889 2.46 60 100 6 20980 1262 10073 43695 2.08

TABLE 14 The effect of oven/dissolution temperature on molecular weightof silk processed under the conditions of 100° C. ExtractionTemperature, 60 min. Extraction Time, and 140° C. Lithium Bromide (LiBr)(Oven/Dissolution Time was varied). Oven Boil Temp Oven Average StdConfidence Time (^(o) C.) Time Mw dev Interval PD 60  60 4 30042 153611183 80705 2.69 60 140 4 15548  7255 33322 2.14

TABLE 15 The effect of oven/dissolution temperature on molecular weightof silk processed under the conditions of 100° C. ExtractionTemperature, 30 min. Extraction Time, and 140° C. Lithium Bromide (LiBr)(Oven/Dissolution Time was varied). Oven Boil Temp Oven Average StdConfidence Time (° C.) Time Mw dev Interval PD 30  60 4 49656  458017306 142478 30 140 4  9025  1102  4493  18127 2.01 30  60 6 59383 1164017641 199889 3.37 30 140 6 13021  5987  28319 2.17

TABLE 16 The effect of oven/dissolution temperature on molecular weightof silk processed under the conditions of 100° C. ExtractionTemperature, 60 min. Extraction Time, and 80° C. Lithium Bromide (LiBr)(Oven/Dissolution Time was varied). Oven Boil Temp Oven Average StdConfidence Time (° C.) Time Mw dev Interval PD 60 60 4 26313  637 1026667442 2.56 60 80 4 30308 4293 12279 74806 2.47 60 60 6 26353 10168 683022.59 60 80 6 25164 238  9637 65706 2.61

In an embodiment, when producing a silk gel, an acid is used to helpfacilitate gelation. In an embodiment, when producing a silk gel thatincludes a neutral or a basic molecule and/or therapeutic agent, an acidcan be added to facilitate gelation. In an embodiment, when producing asilk gel, increasing the pH (making the gel more basic) increases theshelf stability of the gel. In an embodiment, when producing a silk gel,increasing the pH (making the gel more basic) allows for a greaterquantity of an acidic molecule to be loaded into the gel.

In an embodiment, natural additives may be added to the silk gel tofurther stabilize additives. For example, trace elements such asselenium or magnesium or L-methoinine can be used. Further, light-blockcontainers can be added to further increase stability.

In an embodiment, the methods disclosed herein result in a solution withcharacteristics that can be controlled during manufacturing, including,but not limited to: MW—may be varied by changing extraction and/ordissolution time and temp (e.g., LiBr temperature), pressure, andfiltration (e.g., size exclusion chromatography); Structure—removal orcleavage of heavy or light chain of the fibroin protein polymer;Purity—hot water rinse temperature for improved sericin removal orfilter capability for improved particulate removal that adverselyaffects shelf stability of the silk fragment protein mixture solution;Color—the color of the solution can be controlled with, for example,LiBr temp and time; Viscosity; Clarity; and Stability of solution. Theresultant pH of the solution is typically about 7 and can be alteredusing an acid or base as appropriate to storage requirements.

In an embodiment, the above-described SPF mixture solutions may beutilized to coat at least a portion of a fabric which can be used tocreate a textile. In an embodiment, the above-described SPF mixturesolutions may be weaved into yarn that can be used as a fabric in atextile.

FIG. 33 shows two HPLC chromatograms from samples comprising vitamin C.The chromatogram shows peaks from (1) a chemically stabilized sample ofvitamin C at ambient conditions and (2) a sample of vitamin C takenafter 1 hour at ambient conditions without chemical stabilization toprevent oxidation, where degradation products are visible. FIG. 36 is atable summarizing the stability of vitamin C in chemically stabilizedsolutions.

In some embodiments, a composition of the present disclosure can furtherinclude skin penetration enhancers, including, but not limited to,sulfoxides (such as dimethylsulfoxide), pyrrolidones (such as2-pyrrolidone), alcohols (such as ethanol or decanol), azones (such aslaurocapram and 1-dodecylazacycloheptan-2-one), surfactants (includingalkyl carboxylates and their corresponding acids such as oleic acid,fluoroalkylcarboxylates and their corresponding acids, alkyl sulfates,alkyl ether sulfates, docusates such as dioctyl sodium sulfosuccinate,alkyl benzene sulfonates, alkyl ether phosphates, and alkyl aryl etherphosphates), glycols (such as propylene glycol), terpenes (such aslimonene, p-cymene, geraniol, farnesol, eugenol, menthol, terpineol,carveol, carvone, fenchone, and verbenone), and dimethyl isosorbide.

Following are non-limiting examples of suitable ranges for variousparameters in and for preparation of the silk solutions of the presentdisclosure. The silk solutions of the present disclosure may include oneor more, but not necessarily all, of these parameters and may beprepared using various combinations of ranges of such parameters.

In an embodiment, the percent silk in the solution is less than 30%. Inan embodiment, the percent silk in the solution is less than 25%. In anembodiment, the percent silk in the solution is less than 20%. In anembodiment, the percent silk in the solution is less than 19%. In anembodiment, the percent silk in the solution is less than 18%. In anembodiment, the percent silk in the solution is less than 17%. In anembodiment, the percent silk in the solution is less than 16%. In anembodiment, the percent silk in the solution is less than 15%. In anembodiment, the percent silk in the solution is less than 14%. In anembodiment, the percent silk in the solution is less than 13%. In anembodiment, the percent silk in the solution is less than 12%. In anembodiment, the percent silk in the solution is less than 11%. In anembodiment, the percent silk in the solution is less than 10%. In anembodiment, the percent silk in the solution is less than 9%. In anembodiment, the percent silk in the solution is less than 8%. In anembodiment, the percent silk in the solution is less than 7%. In anembodiment, the percent silk in the solution is less than 6%. In anembodiment, the percent silk in the solution is less than 5%. In anembodiment, the percent silk in the solution is less than 4%. In anembodiment, the percent silk in the solution is less than 3%. In anembodiment, the percent silk in the solution is less than 2%. In anembodiment, the percent silk in the solution is less than 1%. In anembodiment, the percent silk in the solution is less than 0.9%. In anembodiment, the percent silk in the solution is less than 0.8%. In anembodiment, the percent silk in the solution is less than 0.7%. In anembodiment, the percent silk in the solution is less than 0.6%. In anembodiment, the percent silk in the solution is less than 0.5%. In anembodiment, the percent silk in the solution is less than 0.4%. In anembodiment, the percent silk in the solution is less than 0.3%. In anembodiment, the percent silk in the solution is less than 0.2%. In anembodiment, the percent silk in the solution is less than 0.1%. In anembodiment, the percent silk in the solution is greater than 0.1%. In anembodiment, the percent silk in the solution is greater than 0.2%. In anembodiment, the percent silk in the solution is greater than 0.3%. In anembodiment, the percent silk in the solution is greater than 0.4%. In anembodiment, the percent silk in the solution is greater than 0.5%. In anembodiment, the percent silk in the solution is greater than 0.6%. In anembodiment, the percent silk in the solution is greater than 0.7%. In anembodiment, the percent silk in the solution is greater than 0.8%. In anembodiment, the percent silk in the solution is greater than 0.9%. In anembodiment, the percent silk in the solution is greater than 1%. In anembodiment, the percent silk in the solution is greater than 2%. In anembodiment, the percent silk in the solution is greater than 3%. In anembodiment, the percent silk in the solution is greater than 4%. In anembodiment, the percent silk in the solution is greater than 5%. In anembodiment, the percent silk in the solution is greater than 6%. In anembodiment, the percent silk in the solution is greater than 7%. In anembodiment, the percent silk in the solution is greater than 8%. In anembodiment, the percent silk in the solution is greater than 9%. In anembodiment, the percent silk in the solution is greater than 10%. In anembodiment, the percent silk in the solution is greater than 11%. In anembodiment, the percent silk in the solution is greater than 12%. In anembodiment, the percent silk in the solution is greater than 13%. In anembodiment, the percent silk in the solution is greater than 14%. In anembodiment, the percent silk in the solution is greater than 15%. In anembodiment, the percent silk in the solution is greater than 16%. In anembodiment, the percent silk in the solution is greater than 17%. In anembodiment, the percent silk in the solution is greater than 18%. In anembodiment, the percent silk in the solution is greater than 19%. In anembodiment, the percent silk in the solution is greater than 20%. In anembodiment, the percent silk in the solution is greater than 25%. In anembodiment, the percent silk in the solution is between 0.1% and 30%. Inan embodiment, the percent silk in the solution is between 0.1% and 25%.In an embodiment, the percent silk in the solution is between 0.1% and20%. In an embodiment, the percent silk in the solution is between 0.1%and 15%. In an embodiment, the percent silk in the solution is between0.1% and 10%. In an embodiment, the percent silk in the solution isbetween 0.1% and 9%. In an embodiment, the percent silk in the solutionis between 0.1% and 8%. In an embodiment, the percent silk in thesolution is between 0.1% and 7%. In an embodiment, the percent silk inthe solution is between 0.1% and 6.5%. In an embodiment, the percentsilk in the solution is between 0.1% and 6%. In an embodiment, thepercent silk in the solution is between 0.1% and 5.5%. In an embodiment,the percent silk in the solution is between 0.1% and 5%. In anembodiment, the percent silk in the solution is between 0.1% and 4.5%.In an embodiment, the percent silk in the solution is between 0.1% and4%. In an embodiment, the percent silk in the solution is between 0.1%and 3.5%. In an embodiment, the percent silk in the solution is between0.1% and 3%. In an embodiment, the percent silk in the solution isbetween 0.1% and 2.5%. In an embodiment, the percent silk in thesolution is between 0.1% and 2.0%. In an embodiment, the percent silk inthe solution is between 0.1% and 2.4%. In an embodiment, the percentsilk in the solution is between 0.5% and 5%. In an embodiment, thepercent silk in the solution is between 0.5% and 4.5%. In an embodiment,the percent silk in the solution is between 0.5% and 4%. In anembodiment, the percent silk in the solution is between 0.5% and 3.5%.In an embodiment, the percent silk in the solution is between 0.5% and3%. In an embodiment, the percent silk in the solution is between 0.5%and 2.5%. In an embodiment, the percent silk in the solution is between1 and 4%. In an embodiment, the percent silk in the solution is between1 and 3.5%. In an embodiment, the percent silk in the solution isbetween 1 and 3%. In an embodiment, the percent silk in the solution isbetween 1 and 2.5%. In an embodiment, the percent silk in the solutionis between 1 and 2.4%. In an embodiment, the percent silk in thesolution is between 1 and 2%. In an embodiment, the percent silk in thesolution is between 20% and 30%. In an embodiment, the percent silk inthe solution is between 0.1% and 6%. In an embodiment, the percent silkin the solution is between 6% and 10%. In an embodiment, the percentsilk in the solution is between 6% and 8%. In an embodiment, the percentsilk in the solution is between 6% and 9%. In an embodiment, the percentsilk in the solution is between 10% and 20%. In an embodiment, thepercent silk in the solution is between 11% and 19%. In an embodiment,the percent silk in the solution is between 12% and 18%. In anembodiment, the percent silk in the solution is between 13% and 17%. Inan embodiment, the percent silk in the solution is between 14% and 16%.In an embodiment, the percent silk in the solution is 2.4%. In anembodiment, the percent silk in the solution is 2.0%.

In an embodiment, the percent sericin in the solution is non-detectableto 30%. In an embodiment, the percent sericin in the solution isnon-detectable to 5%. In an embodiment, the percent sericin in thesolution is 1%. In an embodiment, the percent sericin in the solution is2%. In an embodiment, the percent sericin in the solution is 3%. In anembodiment, the percent sericin in the solution is 4%. In an embodiment,the percent sericin in the solution is 5%. In an embodiment, the percentsericin in the solution is 10%. In an embodiment, the percent sericin inthe solution is 30%.

In an embodiment, the stability of the LiBr-silk fragment solution is 0to 1 year. In an embodiment, the stability of the LiBr-silk fragmentsolution is 0 to 2 years. In an embodiment, the stability of theLiBr-silk fragment solution is 0 to 3 years. In an embodiment, thestability of the LiBr-silk fragment solution is 0 to 4 years. In anembodiment, the stability of the LiBr-silk fragment solution is 0 to 5years. In an embodiment, the stability of the LiBr-silk fragmentsolution is 1 to 2 years. In an embodiment, the stability of theLiBr-silk fragment solution is 1 to 3 years. In an embodiment, thestability of the LiBr-silk fragment solution is 1 to 4 years. In anembodiment, the stability of the LiBr-silk fragment solution is 1 to 5years. In an embodiment, the stability of the LiBr-silk fragmentsolution is 2 to 3 years. In an embodiment, the stability of theLiBr-silk fragment solution is 2 to 4 years. In an embodiment, thestability of the LiBr-silk fragment solution is 2 to 5 years. In anembodiment, the stability of the LiBr-silk fragment solution is 3 to 4years. In an embodiment, the stability of the LiBr-silk fragmentsolution is 3 to 5 years. In an embodiment, the stability of theLiBr-silk fragment solution is 4 to 5 years.

In an embodiment, the stability of a composition of the presentdisclosure is 10 days to 6 months. In an embodiment, the stability of acomposition of the present disclosure is 6 months to 12 months. In anembodiment, the stability of a composition of the present disclosure is12 months to 18 months. In an embodiment, the stability of a compositionof the present disclosure is 18 months to 24 months. In an embodiment,the stability of a composition of the present disclosure is 24 months to30 months. In an embodiment, the stability of a composition of thepresent disclosure is 30 months to 36 months. In an embodiment, thestability of a composition of the present disclosure is 36 months to 48months. In an embodiment, the stability of a composition of the presentdisclosure is 48 months to 60 months.

In an embodiment, a composition of the present disclosure includes puresilk fibroin-based protein fragments having an average weight averagemolecular weight ranging from 6 kDa to 16 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 17 kDa to 38 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 39 kDa to80 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 1 to 5 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 5 to 10 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 10 to 15 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 15 to 20kDa. In an embodiment, a composition of the present disclosure includespure silk fibroin-based protein fragments having an average weightaverage molecular weight ranging from 20 to 25 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 25 to 30 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 30 to 35kDa. In an embodiment, a composition of the present disclosure includespure silk fibroin-based protein fragments having an average weightaverage molecular weight ranging from 35 to 40 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 40 to 45 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 45 to 50kDa. In an embodiment, a composition of the present disclosure includespure silk fibroin-based protein fragments having an average weightaverage molecular weight ranging from 50 to 55 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 55 to 60 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 60 to 65kDa. In an embodiment, a composition of the present disclosure includespure silk fibroin-based protein fragments having an average weightaverage molecular weight ranging from 65 to 70 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 70 to 75 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 75 to 80kDa. In an embodiment, a composition of the present disclosure includespure silk fibroin-based protein fragments having an average weightaverage molecular weight ranging from 80 to 85 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 85 to 90 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 90 to 95kDa. In an embodiment, a composition of the present disclosure includespure silk fibroin-based protein fragments having an average weightaverage molecular weight ranging from 95 to 100 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 100 to 105 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 105 to110 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 110 to 115 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 115 to 120 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 120 to 125 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 125 to130 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 130 to 135 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 135 to 140 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 140 to 145 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 145 to150 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 150 to 155 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 155 to 160 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 160 to 165 kDa. I In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 165 to170 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 170 to 175 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 175 to 180 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 180 to 185 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 185 to190 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 190 to 195 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 195 to 200 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 200 to 205 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 205 to210 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 210 to 215 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 215 to 220 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 220 to 225 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 225 to230 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 230 to 235 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 235 to 240 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 240 to 245 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 245 to250 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 250 to 255 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 255 to 260 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 260 to 265 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 265 to270 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 270 to 275 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 275 to 280 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 280 to 285 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 285 to290 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 290 to 295 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 295 to 300 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 300 to 305 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 305 to310 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 310 to 315 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 315 to 320 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 320 to 325 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 325 to330 kDa. In an embodiment, a composition of the present disclosureincludes pure silk fibroin-based protein fragments having an averageweight average molecular weight ranging from 330 to 335 kDa. In anembodiment, a composition of the present disclosure includes pure silkfibroin-based protein fragments having an average weight averagemolecular weight ranging from 35 to 340 kDa. In an embodiment, acomposition of the present disclosure includes pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from 340 to 345 kDa. In an embodiment, a composition of thepresent disclosure includes pure silk fibroin-based protein fragmentshaving an average weight average molecular weight ranging from 345 to350 kDa.

In an embodiment, a composition of the present disclosure having puresilk fibroin-based protein fragments has a polydispersity ranging fromabout 1 to about 5.0. In an embodiment, a composition of the presentdisclosure having pure silk fibroin-based protein fragments has apolydispersity ranging from about 1.5 to about 3.0. In an embodiment, acomposition of the present disclosure having pure silk fibroin-basedprotein fragments has a polydispersity ranging from about 1 to about1.5. In an embodiment, a composition of the present disclosure havingpure silk fibroin-based protein fragments has a polydispersity rangingfrom about 1.5 to about 2.0. In an embodiment, a composition of thepresent disclosure having pure silk fibroin-based protein fragments hasa polydispersity ranging from about 2.0 to about 2.5. In an embodiment,a composition of the present disclosure having pure silk fibroin-basedprotein fragments, has a polydispersity ranging from about is 2.0 toabout 3.0. In an embodiment, a composition of the present disclosurehaving pure silk fibroin-based protein fragments, has a polydispersityranging from about is 2.5 to about 3.0.

In an embodiment, a composition of the present disclosure having puresilk fibroin-based protein fragments has non-detectable levels of LiBrresiduals. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is between 10 ppm and 1000 ppm. Inan embodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is between 10 ppm and 300 ppm. In an embodiment, theamount of the LiBr residuals in a composition of the present disclosureis less than 25 ppm. In an embodiment, the amount of the LiBr residualsin a composition of the present disclosure is less than 50 ppm. In anembodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is less than 75 ppm. In an embodiment, the amount ofthe LiBr residuals in a composition of the present disclosure is lessthan 100 ppm. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is less than 200 ppm. In anembodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is less than 300 ppm. In an embodiment, the amount ofthe LiBr residuals in a composition of the present disclosure is lessthan 400 ppm. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is less than 500 ppm. In anembodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is less than 600 ppm. In an embodiment, the amount ofthe LiBr residuals in a composition of the present disclosure is lessthan 700 ppm. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is less than 800 ppm. In anembodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is less than 900 ppm. In an embodiment, the amount ofthe LiBr residuals in a composition of the present disclosure is lessthan 1000 ppm. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is non-detectable to 500 ppm. Inan embodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is non-detectable to 450 ppm. In an embodiment, theamount of the LiBr residuals in a composition of the present disclosureis non-detectable to 400 ppm. In an embodiment, the amount of the LiBrresiduals in a composition of the present disclosure is non-detectableto 350 ppm. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is non-detectable to 300 ppm. Inan embodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is non-detectable to 250 ppm. In an embodiment, theamount of the LiBr residuals in a composition of the present disclosureis non-detectable to 200 ppm. In an embodiment, the amount of the LiBrresiduals in a composition of the present disclosure is non-detectableto 150 ppm. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is non-detectable to 100 ppm. Inan embodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is 100 ppm to 200 ppm. In an embodiment, the amountof the LiBr residuals in a composition of the present disclosure is 200ppm to 300 ppm. In an embodiment, the amount of the LiBr residuals in acomposition of the present disclosure is 300 ppm to 400 ppm. In anembodiment, the amount of the LiBr residuals in a composition of thepresent disclosure is 400 ppm to 500 ppm.

In an embodiment, a composition of the present disclosure having puresilk fibroin-based protein fragments, has non-detectable levels ofNa₂CO₃ residuals. In an embodiment, the amount of the Na₂CO₃ residualsin a composition of the present disclosure is less than 100 ppm. In anembodiment, the amount of the Na₂CO₃ residuals in a composition of thepresent disclosure is less than 200 ppm. In an embodiment, the amount ofthe Na₂CO₃ residuals in a composition of the present disclosure is lessthan 300 ppm. In an embodiment, the amount of the Na₂CO₃ residuals in acomposition of the present disclosure is less than 400 ppm. In anembodiment, the amount of the Na₂CO₃ residuals in a composition of thepresent disclosure is less than 500 ppm. In an embodiment, the amount ofthe Na₂CO₃ residuals in a composition of the present disclosure is lessthan 600 ppm. In an embodiment, the amount of the Na₂CO₃ residuals in acomposition of the present disclosure is less than 700 ppm. In anembodiment, the amount of the Na₂CO₃ residuals in a composition of thepresent disclosure is less than 800 ppm. In an embodiment, the amount ofthe Na₂CO₃ residuals in a composition of the present disclosure is lessthan 900 ppm. In an embodiment, the amount of the Na₂CO₃ residuals in acomposition of the present disclosure is less than 1000 ppm. In anembodiment, the amount of the Na₂CO₃ residuals in a composition of thepresent disclosure is non-detectable to 500 ppm. In an embodiment, theamount of the Na₂CO₃ residuals in a composition of the presentdisclosure is non-detectable to 450 ppm. In an embodiment, the amount ofthe Na₂CO₃ residuals in a composition of the present disclosure isnon-detectable to 400 ppm. In an embodiment, the amount of the Na₂CO₃residuals in a composition of the present disclosure is non-detectableto 350 ppm. In an embodiment, the amount of the Na₂CO₃ residuals in acomposition of the present disclosure is non-detectable to 300 ppm. Inan embodiment, the amount of the Na₂CO₃ residuals in a composition ofthe present disclosure is non-detectable to 250 ppm. In an embodiment,the amount of the Na₂CO₃ residuals in a composition of the presentdisclosure is non-detectable to 200 ppm. In an embodiment, the amount ofthe Na₂CO₃ residuals in a composition of the present disclosure isnon-detectable to 150 ppm. In an embodiment, the amount of the Na₂CO₃residuals in a composition of the present disclosure is non-detectableto 100 ppm. In an embodiment, the amount of the Na₂CO₃ residuals in acomposition of the present disclosure is 100 ppm to 200 ppm. In anembodiment, the amount of the Na₂CO₃ residuals in a composition of thepresent disclosure is 200 ppm to 300 ppm. In an embodiment, the amountof the Na₂CO₃ residuals in a composition of the present disclosure is300 ppm to 400 ppm. In an embodiment, the amount of the Na₂CO₃ residualsin a composition of the present disclosure is 400 ppm to 500 ppm.

In an embodiment, the water solubility of pure silk fibroin-basedprotein fragments of the present disclosure is 50 to 100%. In anembodiment, the water solubility of pure silk fibroin-based proteinfragments of the present disclosure is 60 to 100%. In an embodiment, thewater solubility of pure silk fibroin-based protein fragments of thepresent disclosure is 70 to 100%. In an embodiment, the water solubilityof pure silk fibroin-based protein fragments of the present disclosureis 80 to 100%. In an embodiment, the water solubility is 90 to 100%. Inan embodiment, the silk fibroin-based fragments of the presentdisclosure are non-soluble in aqueous solutions.

In an embodiment, the solubility of pure silk fibroin-based proteinfragments of the present disclosure in organic solutions is 50 to 100%.In an embodiment, the solubility of pure silk fibroin-based proteinfragments of the present disclosure in organic solutions is 60 to 100%.In an embodiment, the solubility of pure silk fibroin-based proteinfragments of the present disclosure in organic solutions is 70 to 100%.In an embodiment, the solubility of pure silk fibroin-based proteinfragments of the present disclosure in organic solutions is 80 to 100%.In an embodiment, the solubility of pure silk fibroin-based proteinfragments of the present disclosure in organic solutions is 90 to 100%.In an embodiment, the silk fibroin-based fragments of the presentdisclosure are non-soluble in organic solutions.

In an embodiment, the extraction temperature during a method ofpreparing a composition of the present disclosure is greater than 84° C.In an embodiment, the extraction temperature during a method ofpreparing a composition of the present disclosure is less than 100° C.In an embodiment, the extraction temperature during a method ofpreparing a composition of the present disclosure is 84° C. to 100° C.In an embodiment, the extraction temperature during a method ofpreparing a composition of the present disclosure is 84° C. to 94° C. Inan embodiment, the extraction temperature during a method of preparing acomposition of the present disclosure is 94° C. to 100° C.

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the described embodiments, and are not intended to limitthe scope of what the inventors regard as their invention nor are theyintended to represent that the experiments below are all or the onlyexperiments performed. Efforts have been made to ensure accuracy withrespect to numbers used (e.g. amounts, temperature, etc.) but someexperimental errors and deviations should be accounted for. Unlessindicated otherwise, parts are parts by weight, molecular weight isweight average molecular weight, temperature is in degrees Centigrade,and pressure is at or near atmospheric.

There is disclosed a textile that is at least partially surface treatedwith an aqueous solution of pure silk fibroin-based protein fragments ofthe present disclosure so as to result in a silk coating on the textile.In an embodiment, the silk coating of the present disclosure isavailable in a spray can and can be sprayed on any textile by aconsumer. In an embodiment, a textile comprising a silk coating of thepresent disclosure is sold to a consumer. In an embodiment, a textile ofthe present disclosure is used in constructing actionsportswear/apparel. In an embodiment, a silk coating of the presentdisclosure is positioned on the underlining of apparel. In anembodiment, a silk coating of the present disclosure is positioned onthe shell, the lining, or the interlining of apparel. In an embodiment,apparel is partially made from a silk coated textile of the presentdisclosure and partially made from an uncoated textile. In anembodiment, apparel partially made from a silk coated textile andpartially made from an uncoated textile combines an uncoated inertsynthetic material with a silk coated inert synthetic material. Examplesof inert synthetic material include, but are not limited to, polyester,polyamide, polyaramid, polytetrafluorethylene, polyethylene,polypropylene, polyurethane, silicone, mixtures of polyurethane andpolyethylenglycol, ultrahigh molecular weight polyethylene,high-performance polyethylene, and mixtures thereof. In an embodiment,apparel partially made from a silk coated textile and partially madefrom an uncoated textile combines an elastomeric material at leastpartially covered with a silk coating of the present disclosure. In anembodiment, the percentage of silk to elastomeric material can be variedto achieve desired shrink or wrinkle resistant properties.

In an embodiment, a silk coating of the present disclosure is visible.In an embodiment, a silk coating of the present disclosure positioned onapparel helps control skin temperature. In an embodiment, a silk coatingof the present disclosure positioned on apparel helps control fluidtransfer away from the skin. In an embodiment, a silk coating of thepresent disclosure positioned on apparel has a soft feel against theskin decreasing abrasions from fabric on skin. In an embodiment, a silkcoating of the present disclosure positioned on a textile has propertiesthat confer at least one of wrinkle resistance, shrinkage resistance, ormachine washability to the textile. In an embodiment, a silk coatedtextile of the present disclosure is 100% machine washable and drycleanable. In an embodiment, a silk coated textile of the presentdisclosure is 100% waterproof. In an embodiment, a silk coated textileof the present disclosure is wrinkle resistant. In an embodiment, a silkcoated textile of the present disclosure is shrink resistant. In anembodiment, a silk coated textile of the present disclosure has thequalities of being waterproof, breathable, and elastic and possess anumber of other qualities which are highly desirable in actionsportswear. In an embodiment, a silk coated textile of the presentdisclosure manufactured from a silk fabric of the present disclosurefurther includes LYCRA® brand spandex fibers.

In an embodiment, a textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure is a breathable fabric. In an embodiment, a textile at leastpartially coated with an aqueous solution of pure silk fibroin-basedprotein fragments of the present disclosure is a water-resistant fabric.In an embodiment, a textile at least partially coated with an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure is a shrink-resistant fabric. In an embodiment, a textile atleast partially coated with an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure is amachine-washable fabric. In an embodiment, a textile at least partiallycoated with an aqueous solution of pure silk fibroin-based proteinfragments of the present disclosure is a wrinkle resistant fabric. In anembodiment, textile at least partially coated with an aqueous solutionof pure silk fibroin-based protein fragments of the present disclosureprovides moisture and vitamins to the skin.

In an embodiment, an aqueous solution of pure silk fibroin-based proteinfragments of the present disclosure is used to coat a textile. In anembodiment, the concentration of silk in the solution ranges from about0.1% to about 20.0%. In an embodiment, the concentration of silk in thesolution ranges from about 0.1% to about 15.0%. In an embodiment, theconcentration of silk in the solution ranges from about 0.5% to about10.0%. In an embodiment, the concentration of silk in the solutionranges from about 1.0% to about 5.0%. In an embodiment, an aqueoussolution of pure silk fibroin-based protein fragments of the presentdisclosure is applied directly to a fabric. Alternatively, silkmicrosphere and any additives may be used for coating a fabric. In anembodiment, additives can be added to an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure before coating(e.g., alcohols) to further enhance material properties. In anembodiment, a silk coating of the present disclosure can have a patternto optimize properties of the silk on the fabric. In an embodiment, acoating is applied to a fabric under tension and/or lax to varypenetration in to the fabric.

In an embodiment, a silk coating of the present disclosure can beapplied at the yarn level, followed by creation of a fabric once theyarn is coated. In an embodiment, an aqueous solution of pure silkfibroin-based protein fragments of the present disclosure can be spuninto fibers to make a silk fabric and/or silk fabric blend with othermaterials known in the apparel industry.

In an embodiment, a method for silk coating a fabric includes immersionof the fabric in any of the aqueous solutions of pure silk fibroin-basedprotein fragments of the present disclosure. In an embodiment, a methodfor silk coating a fabric includes spraying. In an embodiment, a methodfor silk coating a fabric includes chemical vapor deposition. In anembodiment, a method for silk coating a fabric includes electrochemicalcoating. In an embodiment, a method for silk coating a fabric includesknife coating to spread any of the aqueous solutions of pure silkfibroin-based protein fragments of the present disclosure onto thefabric. The coated fabric may then be air dried, dried under heat/airflow, or cross-linked to the fabric surface. In an embodiment, a dryingprocess includes curing with additives and/or ambient condition.

EXAMPLES Example 1. Tangential Flow Filtration (TFF) to Remove Solventfrom Dissolved Silk Solutions

A variety of % silk concentrations have been produced through the use ofTangential Flow Filtration (TFF). In all cases a 1% silk solution wasused as the input feed. A range of 750-18,000 mL of 1% silk solution wasused as the starting volume. Solution is diafiltered in the TFF toremove lithium bromide. Once below a specified level of residual LiBr,solution undergoes ultrafiltration to increase the concentration throughremoval of water. See examples below.

7.30% Silk Solution: A 7.30% silk solution was produced beginning with30 minute extraction batches of 100 g silk cocoons per batch. Extractedsilk fibers were then dissolved using 100 C 9.3M LiBr in a 100 C ovenfor 1 hour. 100 g of silk fibers were dissolved per batch to create 20%silk in LiBr. Dissolved silk in LiBr was then diluted to 1% silk andfiltered through a 5 um filter to remove large debris. 15,500 mL of 1%,filtered silk solution was used as the starting volume/diafiltrationvolume for TFF. Once LiBr was removed, the solution was ultrafiltered toa volume around 1300 mL. 1262 mL of 7.30% silk was then collected. Waterwas added to the feed to help remove the remaining solution and 547 mLof 3.91% silk was then collected.

6.44% Silk Solution: A 6.44% silk solution was produced beginning with60 minute extraction batches of a mix of 25, 33, 50, 75 and 100 g silkcocoons per batch. Extracted silk fibers were then dissolved using 100 C9.3M LiBr in a 100 C oven for 1 hour. 35, 42, 50 and 71 g per batch ofsilk fibers were dissolved to create 20% silk in LiBr and combined.Dissolved silk in LiBr was then diluted to 1% silk and filtered througha 5 um filter to remove large debris. 17,000 mL of 1%, filtered silksolution was used as the starting volume/diafiltration volume for TFF.Once LiBr was removed, the solution was ultrafiltered to a volume around3000 mL. 1490 mL of 6.44% silk was then collected. Water was added tothe feed to help remove the remaining solution and 1454 mL of 4.88% silkwas then collected

2.70% Silk Solution: A 2.70% silk solution was produced beginning with60 minute extraction batches of 25 g silk cocoons per batch. Extractedsilk fibers were then dissolved using 100 C 9.3M LiBr in a 100 C ovenfor 1 hour. 35.48 g of silk fibers were dissolved per batch to create20% silk in LiBr. Dissolved silk in LiBr was then diluted to 1% silk andfiltered through a 5 um filter to remove large debris. 1000 mL of 1%,filtered silk solution was used as the starting volume/diafiltrationvolume for TFF. Once LiBr was removed, the solution was ultrafiltered toa volume around 300 mL. 312 mL of 2.7% silk was then collected.

Example 2. Preparation of Silk Gels

TABLE 17 Gel Samples—Silk gel formulations including additives,concentration of silk and additive, gelation conditions and gelationtimes. Sample mL 2% silk Mass Vit C Ratio silk: Amount of Temp/ Days toName solution (g) VitC Additive additive Treatment Gelation 1 10 0.045:01 None None RT 8 2 10 0.08 2.5:1 None None RT 8 3 10 0.2 1:01 NoneNone RT 8 4 10 0.4 1:02 None None RT 14 5 10 0.8 1:04 None None RT None6 10 0.04 5:01 None None Fridge ~39 7 10 0.08 2.5:1 None None Fridge ~398 10 0.2 1:01 None None Fridge ~39 9 10 0.4 1:02 None None Fridge None10 10 0.8 1:04 None None Fridge None 11 10 0.2 1:01 None None RT/Shake 8vigorously O-1 10 0.04 5:01 None None 37 C. Oven 3 O-2 10 0.04 5:01 NoneNone 50 C. Oven 2 O-3 10 0.2 1:01 None None 37 C. Oven 4 O-4 10 0.2 1:01None None 50 C. Oven 3 M 40 0.16 5:01 None None RT 5 D 40 0.16 5:01 NoneNone RT 5 E1 10 0.04 5:01 VitE 1 drop RT 7 E2 10 0.04 5:01 VitE 3 dropsRT 7 E3 10 0 None VitE 1 drop RT None E4 10 0 None VitE 3 drops RT NoneL1 10 0.04 5:01 Lemon  300 uL RT 6 L2 10 0.04 5:01 Lemon Juice  300 uLRT 6 L3 10 0.04 5:01 Lemon Juice 1000 uL RT 5 L4 10 0 None Lemon  300 uLRT 6 L5 10 0 None Lemon Juice  300 uL RT 7 Jar 1 20 0.08 5:01 LemonJuice 2000 uL RT 5-7 Jar 2 5 0.02 5:01 Lemongrass 1 drop RT 2-3 Oil R-110 0.04 5:01 Rosemary 1 drop RT 7 Oil T-1 10 0.04 5:01 None None RT/Tube7 RO-1 10 0.04 5:01 Rose Oil 1 drop RT 6 RO-2 10 None None Rose Oil 1drop RT None

Ratio of Silk to Vitamin C

Samples 1-10 were used to examine the effect of silk to vitamin C ratioon serum gelation. Samples 1-3 with less vitamin C gelled quicker thansamples 4 and 5. All other conditions were kept constant. Samples 6-8with less vitamin C gelled quicker than samples 9 and 10. All otherconditions were kept constant. It is concluded that decreasing the ratioof silk to vitamin C (increasing the amount of vitamin C), will lengthenthe time to gel creation. At ratios with small amounts of vitamin C,days to gel creation did not vary greatly.

Physical Stimulation

Samples 3 and 11 were used to examine the effect of physical stimulationon serum gelation. Each sample was prepared under the same conditions.Sample 11 was vigorously shaken for about 3 minutes after addition ofvitamin C. Treatment of 3 and 11 was otherwise the same. The shakingresulted in bubbles but did not significantly change gel creation time.

Temperature Treatment

Samples 1, 3, 6, 8, O-1, O-2, O-3, and O-4 were used to examine theeffect of temperature treatment on serum gelation time. Samples 1, 6,O-1, and O-2 were identical other than temperature treatment. Samples 3,8, O-3, and O-4 were identical other than temperature treatment. The twogroups differed in silk to vitamin C ratio. Time to serum gelation wasdirectly related to temperature treatment with a higher temperatureresulting in quicker serum gelation.

Solution Volume

Samples 1, M and D were used to examine the effect of solution volume onserum gelation time. Samples M and D varied from sample 1 only by anincreased solution volume. Samples M and D gelled in 5 days while sample1 gelled in 8 days. Samples M and D were definitively noticed to begelled on the day of gelling while sample 1 gelled over a weekend.

Additives

Samples E1, E2, E3, E4, L1, L2, L3, L4, L5, Jar 2, R1, RO-1 and RO-2were used to examine the effect of additives on serum gelation time.Samples E1-4 contained Vitamin E. Only samples E1 and E2 containedvitamin C and only these two samples gelled. Vitamin E can be added to asolution to become a gel but it appears that another additive may beneeded to create a gel. Samples L1-5 contained a form of lemon juice.Samples L1 and L4 had juice directly from a lemon while samples L2, L3and L5 contained lemon juice from a plastic lemon container. Samples L4and L5 did not have vitamin C while all others did. All samples gelledshowing that lemon juice can create gel on its own. Amount of lemonjuice and type of lemon juice had little effect on gelation time. SampleJar 2 contained lemon grass oil which formed an albumen like substancewhen initially added. This sample also had vitamin C but gelation timewas significantly quicker than with other vitamin C samples. Sample R1contained rosemary oil, which seemed to be soluble, as well as vitaminC. The sample gelled in a similar time frame to other samples with onlyvitamin C. Samples RO-1 and RO-2 contained rose oil while only RO-1 hadvitamin C. Only RO-1 gelled showing that rose oil will not create a gelquickly on its own. In both cases the rose oil was immiscible andvisible as yellow bubbles.

Aqueous silk fibroin-based fragment solution and essential oils areimmiscible liquids. In an embodiment, to increase the fragrance of thesilk fibroin-based fragment solution, without entrapping oils within thesolution, the solution is mixed with the essential oil with the use of astir bar. The stir bar is rotated at a speed such that some turbulenceis observed in the mixture, thus causing contact between the fragrantessential oil and the molecules in solution, adding a scent to thesolution. Before casting of product from the solution, mixing may bestopped and the oil allowed to separate to the top of the solution.Dispensing from the bottom fraction of the solution into the finalproduct allows for fragrance without visible essential oil within thefinal product.

Alternatively, the silk fibroin-based solution and essential oil can becombined with or without additional ingredients and/or an emulsifier tocreate a composition containing both ingredients.

In an embodiment, mixing of the solution as described above can reducegelation time if the solution is used to create a gel formulation.

Vessel

Samples T1 and Jar 1 were used to examine the effect of casting vesselon serum gelation time. Jar 1 was cast in a glass jar while T1 was castin an aluminum tube. Both samples gelled and did not affect serum geltime.

Summary

All treatments of silk solution for gel solution were in a conical tubeat room temperature unless otherwise stated. The ratio of silk tovitamin C did affect the ability of a solution to gel as ratios above1:2 did not gel and a 1:2 ratio took twice as long as other lower ratios(5:1, 2.5:1, 1:1). Temperature affected gel creation time with highertemperatures resulting in quicker gel times. 50° C. treatment gelled inas quick as 2 days, 37° C. treatment gelled in as quick as 3 days, roomtemperature treatment gelled in 5-8 days and storage in a refrigeratortook at least 39 days to gel. The effects of additives on gel creationwere dependent on the additive. Vitamin E, Rosemary Oil and Rose Oil allhad no effect on gel creation. Each of these additives did not preventgelation or affect the time to gelation. Each also required the presenceof vitamin C to gel. Lemon juice from a fresh lemon, pre-squeezed lemonjuice from a plastic lemon container and lemon grass oil did affect gelcreation. Without wishing to be bound by theory, it is believed that thelower pH as a result of these additives is the reason the additives hadan impact on decreasing gelation time. Both lemon juice types were ableto cause gelation without the presence of vitamin C. This occurred inthe same number of days as with vitamin C. The lemongrass oil was ableto decrease the number of days to gelation to 2-3 days. All additivesappeared soluble other than lemongrass oil and rose oil. Rose oilremained in yellow bubbles while the lemongrass oil was partiallysoluble and formed an albumen like chunk. In an embodiment, oils thatare not fully soluble, can still be suspended within the gel as anadditive. Physical stimulation by shaking, vessel the solution was castinto and solution volume did not affect gelation time.

TABLE 18 Concentration of vitamin C in various gel formulations. SampleConcentration of Weight Vitamin C (mg/g) Sample Info (mg) In SampleAverage Rosemary 685.7 3.2511 3.2657 (Room 3.2804 Temperature 638 3.33363.3334 storage) 3.3332 Lemongrass 646 2.8672 2.877 (Room 2.8868Temperature 645.5 2.9051 2.9051 storage) 2.9052 Rosemary 645.2 3.90633.9147 (Room 3.923 Temperature; 649 3.9443 3.9374 Foil Covered 3.9305storage) Lemongrass 630.1 3.8253 3.8274 (Room 3.8295 Temperature; 660.43.8283 3.8253 Foil Covered 3.8222 storage) Rosemary 672.4 5.1616 5.1484(Fridge, Foil 5.1352 Covered 616.5 5.1984 5.201 storage) 5.2036Lemongrass 640.5 5.1871 5.1824 (Fridge, Foil 5.1776 Covered 627.7 5.20985.2126 storage) 5.2154

Example 3. Preparation of Silk Gels

Additional gels may be prepared according to Table 19, Table 20, Table21, and Table 22.

TABLE 19 Lemongrass Gel % Silk Solution 2% Quantity Vitamin C 100 mg/15mL solution Quantity Lemongrass Oil  20 μL/15 mL solution

TABLE 20 Rosemary Gel % Silk Solution 2% Quantity Vitamin C 100 mg/15 mLsolution Quantity Rosemary Oil  20 μL/50 mL solution

TABLE 21 Lemongrass Gel (50 mL) % Silk Solution (60 minute boil, 25 kDA)2% Quantity Vitamin C (ascorbyl glucoside) 12.82 mg/mL solution (641 mgtotal) Quantity Lemongrass Oil 1.33 μL/mL solution pH 4

TABLE 22 Rosemary Gel (50 mL) % Silk Solution (60 minute boil, 25 kDA)2% Quantity Vitamin C (ascorbyl glucoside) 12.82 mg/mL solution (641 mgtotal) Quantity Rosemary Oil 0.8 μL/mL solution pH 4

Gels of the present disclosure can be made with about 0.5% to about 8%silk solutions. Gels of the present disclosure can be made with ascorbylglucoside at concentrations of about 0.67% to about 15% w/v. Gels of thepresent disclosure be clear/white in color. Gels of the presentdisclosure can have a consistency that is easily spread and absorbed bythe skin. Gels of the present disclosure can produce no visual residueor oily feel after application. Gels of the present disclosure do notbrown over time.

Silk gels with essential oils were prepared by diluting a silk solutionof the present disclosure to 2%. Vitamin C was added to the solution andallowed to dissolve. The essential oil was added, stirred and dissolved.The solution was aliquot into jars.

Example 4. Coating Fabrics with Aqueous Silk Solutions

TABLE 23 Silk Solution Characteristics Molecular Weight: 57 kDaPolydispersity: 1.6 % Silk 5.0% 3.0% 1.0% 0.5% Process ParametersExtraction Boil Time: 30 minutes Boil Temperature: 100° C. RinseTemperature:  60° C. Dissolution LiBr Temperature: 100° C. OvenTemperature: 100° C. Oven Time: 60 minutes

TABLE 24 Silk Solution Characteristics Molecular Weight: 25 kDaPolydispersity: 2.4 % Silk 5.0% 3.0% 1.0% 0.5% Process ParametersExtraction Boil Time: 60 minutes Boil Temperature: 100° C. RinseTemperature:  60° C. Dissolution LiBr Temperature: 100° C. OvenTemperature: 100° C. Oven Time: 60 minutes

Silk Solution and Silk Gel Application to Fabric and Yarn Samples

Three 50 mm diameter fabric samples from each of three different fabricmaterials, cotton, polyester, and nylon/LYCRA®, were placed in plasticcontainers. about 0.3 mL of about 5.8% silk fibroin solution wasdeposited using a 1 mL syringe and 18 gauge needle on two samples ofeach material, and allowed to sit for about 1 minute. About 0.3 mL ofdenatured alcohol (containing methanol and ethanol) was then depositedusing a 1 mL syringe and 30 gauge needle on one of the silk-coatedsamples of each material.

In an additional experiment, silk gel with Rosemary Essential Oil(water, silk, ascorbyl glucoside, rosemary essential oil) was collectedon a tip and applied to half the length of 2 pieces of 400 um tencelyarn. One sample was then wetted with about 0.3 mL alcohol.

Silk Solution Dip Test

Polyester fabric samples were dipped in silk fibroin solutions ofvarying concentration. Samples were placed in incubator with air flow onfoil and allowed to dry at about 22.5° C. for about 15.5 hours. Changein mass before and after silk coating was measured.

TABLE 25 Polyester Fabric Samples with Silk Coatings of the PresentDisclosure Silk Fibroin Mass after Concentration Starting Mass coatingChange Average (%) (g) (g) (%) Change (%) 1 0.25 0.26 +4 −3% 0.30 0.27−10 0.24 0.24 0 0.22 0.21 −5 3 0.30 0.36 +20 15% 0.28 0.31 +11 0.29 0.33+14 0.29 0.34 +15 5 0.25 0.29 +16 16% 0.28 0.33 +18 0.31 0.35 +13 0.270.31 +15

Silk Solution Spray Test

A spray test was performed to verify the handle impact of silk fibroinsolution sprayed on polyester fabric. About 0.5% silk fibroin solutionwas applied to a 4 inch by 4 inch square of polyester fabric using aspray gun from a distance of about 10 inches. Three passes werecompleted from left to right and from right to left (six passes total).Samples were placed in a 50° C. oven on aluminum foil over a water bathfor about 1.5 hours. Methods were repeated with a second polyesterfabric sample with an about 5.8% silk fibroin solution sprayapplication. No change in material hand was observed in samples sprayedwith either 0.5% or 5.8% solutions. Perceived increase in materialssmoothness was observed for samples sprayed with either the 0.5% and5.8% solutions.

Example 5. Optimized Fabric Coating Processes

The coating processes described in Table 26 were used to producemultiple fabric samples for performance testing, as described in moredetail below.

TABLE 26 Coating Processes. 1 Spray 1.1 Material for coating 1.1.1 corkboard 24” × 36” Hobby Lobby part 132894 1.1.2 Covered the cork boardwith polyester interlock fabric 1.1.3 Saw horse for support 1.1.4Several clamps for holding cork panel to saw horse 1.1.5 Double filterto remove oil residue from compressor and dehumidificaton salt 1.1.6Iwata eclipse MP-CS airbrush 1.1.7 Husky 30.3 liter tank compressionsystem 1.1.8 Push pin to hold fabric on cork panel Hobby Lobby part#523456 1.2 Material for preparation 1.2.1 Scissor 1.2.2 Ruler 1.2.3Balance AWS model Pnx-203 1.3 Material for drying 1.3.1 Wolf stove setup at 150° F. maintaining 71-78° C. with fan system. 1.3.2 Flat bakingsheet 1.3.3 Aluminum foil 1.3.4 SC 307T thermometer with probe 1.4Execution 1.1.1 lay fabric to be coated on top of cork panel coveredwith polyester fabric 1.1.2 secure fabric with pin to the cork panel1.1.3 set compressor with oil and humidity filters 1.1.4 set airpressure supply to 55 psi 1.1.5 load solution to airbrush gun 1.1.6position airbrush gun approximately 10 inches from board 1.1.7 pull theairbrush gun trigger and over spray 2 inches side to side the fabric tobe coated 1.1.8 remove pin from cork panel and place coated fabric onaluminum foil 1.1.9 place coated fabric in oven for 30-60 min at 150° C.2 Stencil/Spray 2.1 Material for coating 2.1.1 cork board 24” × 36”Hobby Lobby part 132894 2.1.2 Covered the cork board with polyesterinterlock fabric 2.1.3 Saw horse for support 2.1.4 Several clamps forholding cork panel to saw horse 2.1.5 Double filter to remove oilresidue from compressor and dehumidificaton salt 2.1.6 Iwata eclipseMP-CS airbrush 2.1.7 Husky 30.3 liter tank compression system 2.1.8 Pushpin to hold fabric on cork panel Hobby Lobby part #523456 2.1.9 Stencilpattern SKU #75244 Lincaine 12” × 24” × 0.020” Hobby Lobby 2.2 Materialfor preparation 2.2.1 Scissor 2.2.2 Ruler 2.2.3 Balance AWS modelPnx-203 2.3 Material for drying 2.3.1 Wolf stove set up at 150° F.maintaining 71-78° C. with fan system. 2.3.2 Flat baking sheet 2.3.3Aluminum foil 2.3.4 SC 307T thermometer with probe 2.4 Execution 2.4.1lay fabric to be coated on top of cork panel covered with polyesterfabric 2.4.2 lay stencil pattern on top of fabric 2.4.3 secure stencilwith pin to the cork panel 2.4.4 set compressor with oil and humidityfilters 2.4.5 set air pressure supply to 55 psi 2.4.6 load solution toairbrush gun 2.4.7 position airbrush gun approximately 10 inches fromboard 2.4.8 pull the airbrush gun trigger and over spray 2 inches sideto side the fabric to be coated 2.4.9 remove pin from cork panel andplace coated fabric on aluminum foil 2.4.10 place coated fabric in ovenfor 30-60 min at 150° C. 3 Screen print 3.1 Material for coating 3.1.1cork board 24” × 36” Hobby Lobby part 132894 3.1.2 Covered the corkboard with polyester interlock fabric 3.1.3 Saw horse for support 3.1.4Several clamps for holding cork panel to saw horse 3.1.5 screen printframe 12” × 18” part #4710 made by Speed Ball 3.1.6 silicon spatula 3.2Material for preparation 3.2.1 Scissor 3.2.2 Ruler 3.2.3 Balance AWSmodel Pnx-203 3.3 Material for drying 3.3.1 Wolf stove set up at 150° F.maintaining 71-78° C. with fan system. 3.3.2 Flat baking sheet 3.3.3Aluminum foil 3.3.4 SC 307T thermometer with probe 3.4 Execution 3.4.1lay fabric to be coated on top of cork panel covered with polyesterfabric 3.4.2 lay screen print frame on top of fabric 3.4.3 load solutionto one edge of the screen print frame 3.4.4 with a silicon spatula movesolution across the screen print frame until the entire fabric to becoated surface is covered 3.4.5 remove screen print frame and placecoated fabric on aluminum foil 3.4.6 place coated fabric in oven for30-60 min at 150° C. 4 Bath 4.1 Material for coating 4.1.1 cork board24” × 36” Hobby Lobby part 132894 4.1.2 Covered the cork board withpolyester interlock fabric 4.1.3 Saw horse for support 4.1.4 Severalclamps for holding cork panel to saw horse 4.1.5 Paint tray liner Item#: 170418 Model #: LOWES0-PK170418 at Lowes Hardware 4.1.6 Noodle makingmachine Imperia model #15-4590 4.2 Material for preparation 4.2.1Scissor 4.2.2 Ruler 4.2.3 Balance AWS model Pnx-203 4.3 Material fordrying 4.3.1 Wolf stove set up at 150° F. maintaining 71-78° C. with fansystem. 4.3.2 Flat baking sheet 4.3.3 Aluminum foil 4.3.4 SC 307Tthermometer with probe 4.4 Execution 4.4.1 load silk solution inside thepaint tray liner well 4.4.2 immerse the fabric sample to be coatedinside the silk solution until it is all saturated 4.4.3 pass thesaturated fabric between pressure roller (noodle making machine) toremove any excess solution 4.4.4 place coated fabric on aluminum foil4.4.5 place coated fabric in oven for 30-60 min at 150° C.

The products produced using the coating processes described above weretested for accumulative one way transport capability (or index) andother properties using Association of Textile, Apparel & MaterialsProfessionals (AATCC) test method 195-2012 for the measurement,evaluation, and classification of liquid moisture management propertiesof textile fabrics. The details of the test methods are available fromAATCC, and a synopsis of the methods and calculations is provided here.The absorption rate (ART) (top surface) and (ARB) (bottom surface) isdefined as the average speed of liquid moisture absorption for the topand bottom surfaces of the specimen during the initial change of watercontent during a test. The accumulative one-way transport capability (R)is defined as the difference between the area of the liquid moisturecontent curves of the top and bottom surfaces of a specimen with respectto time. The bottom surface (B) is defined for testing purposes as theside of the specimen placed down against the lower electrical sensorwhich is the side of the fabric that would be the outer exposed surfaceof a garment when it is worn or product when it is used. The top surface(T) for testing purposes is defined as the side of a specimen that, whenthe specimen is placed on the lower electrical sensor, is facing theupper sensor. This is the side of the fabric that would come in contactwith the skin when a garment is worn or when a product is used. Themaximum wetted radius (MWRT) and (MWRB) (mm) is defined as the greatestring radius measured on the top and bottom surfaces. Moisture managementis defined, for liquid moisture management testing, as the engineered orinherent transport of aqueous liquids such as perspiration or water(relates to comfort) and includes both liquid and vapor forms of water.The overall (liquid) moisture management capability (OMMC), an index ofthe overall capability of a fabric to transport liquid moisture ascalculated by combining three measured attributes of performance: theliquid moisture absorption rate on the bottom surface (ARB), the one wayliquid transport capability (R), and the maximum liquid moisturespreading speed on the bottom surface (SS_(B)). The spreading speed(SS_(i)) is defined as the accumulated rate of surface wetting from thecenter of the specimen where the test solution is dropped to the maximumwetted radius. The total water content (U) (%) is defined as the sum ofthe percent water content of the top and bottom surfaces. The wettingtime (WTT) (top surface) and (WTB) (bottom surface) is defined as thetime in seconds when the top and bottom surfaces of the specimen beginto be wetted after the test is started.

A moisture management tester (MMT) is used to perform the test. Theaccumulative one way liquid transport capability (R) is calculated as:[Area (U_(B))−Area (UT)]/total testing time. The OMMC is calculated as:OMMC=C₁*AR_(B_ndv)+C₂*R_(nvd)+C3*SS_(B_ndv), where C₁, C₂, and C₃ arethe weighting values * for AR_(B_ndv), R_(ndv) and SS_(B_ndv);(ARB)=absorption rate; (R)=one-way transport capability, and(SS_(B))=spreading speed. Detailed calculations of these parameters, andother parameters of interest, are provided in AATCC test method195-2012.

A description of the samples used is given in Table 27.

TABLE 27 Description of samples. Sample ID Description 15051201 nocoating (polyester) 15051301 1% silk solution spray coating on 1505120115051302 0.1% silk solution spray coating on 15051201 15051303 0.05%silk solution spray coating on 15051201 15051304 1% silk solution spraystencil coating on 15051201 15051305 0.1% silk solution spray stencilcoating on 15051201 15051306 0.05% silk solution spray stencil coatingon 15051201 15051401 1% silk solution bath coating on 15051201 150514020.1% silk solution bath coating on 15051201 15051403 0.05% silk solutionbath coating on 15051201 15051404 PureProC screen print on 1505120115042001 non wicking finished 15042002 semifinished before final setting15042003 with wicking finished 15042101 non wicking finished (15042001)1% silk solution spray coating 15042102 non wicking finished (15042001)0.1% silk solution spray coating 15061206 non wicking finished(15042001) 1% silk solution stencil coating 15061207 non wickingfinished (15042001) 1% silk solution bath coating 15061205 non wickingfinished (15042001) 0.1% silk solution stencil coating 15061209 nonwicking finished (15042001) 0.1% silk solution bath coating 15061201semifinished before final setting (15042002) 1% silk solution spraycoating 15061203 semifinished before final setting (15042002) 1% silksolution stencil coating 15061208 semifinished before final setting(15042002) 1% silk solution bath coating 15061202 semifinished beforefinal setting (15042002) 0.1% silk solution spray coating 15061204semifinished before final setting (15042002) 0.1% silk solution stencilcoating 15061210 semifinished before final setting (15042002) 0.1% silksolution bath coating

The results of the tests are depicted in FIG. 57A through FIG. 86B andillustrate the superior performance of silk coated fabric, includingsuperior performance with respect to accumulative one way transportcapability (index) and overall moisture management capability.

Example 6. Antimicrobial Properties of Silk Coatings on Fabrics

The antimicrobial properties of silk coatings were testing on fourmaterials: a cotton/LYCRA jersey (15051201), a cotton/LYCRA jersey with1% silk fibroin solution (sfs) bath coating (15070701), apolyester/LYCRA finish after final setting (15042003), and apolyester/LYCRA semi-finished 1% sfs bath coating (15070702) (whereinLYCRA is the trade name of a polyester-polyurethane copolymer). AATCCtest method 100-2012 for the assessment of antibacterial finishes ontextile materials was used. The details of the test method are availablefrom AATCC. Briefly, the tests were performed using tryptic soy broth asa growth medium, a sample size of 4 layers, autoclave sterilization, 100mL Letheen broth with Tween for neutralization, a target inoculationlevel of 1-2×10⁵ CFU/mL, 5% nutrient broth as an inoculent carrier anddilution medium, a contact time of 18 to 24 hours, and a temperature of37+/−2° C.

The results of the tests are summarized in Table 28 and are depicted inFIG. 87 to FIG. 92 , and illustrate the superior antimicrobialperformance of the silk-coated fabrics.

TABLE 28 Antimicrobial test results. Results: cfu/sample Zero Contact 24hr Contact Percent sample # bacteria Time Time Reduction 15051201Staphylococcus aureus ATCC 6538 1.23E+05 4.90E+06 −3883.74% Klebsiellapneumoniae ATCC 4352 1.65E+05 4.90E+06 −2869.70% 15070701 Staphylococcusaureus ATCC 6538 1.23E+05 4.90E+06 −3883.74% Klebsiella pneumoniae ATCC4352 1.65E+05 4.90E+06 −2869.70% 15042003 Staphylococcus aureus ATCC6538 1.23E+05 4.90E+06 −3883.74% Klebsiella pneumoniae ATCC 43521.65E+05 4.90E+06 −2869.70% 15070702 Staphylococcus aureus ATCC 65381.23E+05 1.03E+04   91.63% Klebsiella pneumoniae ATCC 4352 1.65E+052.33E+05  −40.91%

Example 7. Methods of Preparing Fabrics with Silk Coatings

A method for preparing an aqueous solution of pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from about 6 kDa to about 16 kDa includes the steps of:degumming a silk source by adding the silk source to a boiling (100° C.)aqueous solution of sodium carbonate for a treatment time of betweenabout 30 minutes to about 60 minutes; removing sericin from the solutionto produce a silk fibroin extract comprising non-detectable levels ofsericin; draining the solution from the silk fibroin extract; dissolvingthe silk fibroin extract in a solution of lithium bromide having astarting temperature upon placement of the silk fibroin extract in thelithium bromide solution that ranges from about 60° C. to about 140° C.;maintaining the solution of silk fibroin-lithium bromide in an ovenhaving a temperature of about 140° C. for a period of at least 1 hour;removing the lithium bromide from the silk fibroin extract; andproducing an aqueous solution of silk protein fragments, the aqueoussolution comprising: fragments having an average weight averagemolecular weight ranging from about 6 kDa to about 16 kDa, and whereinthe aqueous solution of pure silk fibroin-based protein fragmentscomprises a polydispersity of between about 1.5 and about 3.0. Themethod may further comprise drying the silk fibroin extract prior to thedissolving step. The aqueous solution of pure silk fibroin-based proteinfragments may comprise lithium bromide residuals of less than 300 ppm asmeasured using a high-performance liquid chromatography lithium bromideassay. The aqueous solution of pure silk fibroin-based protein fragmentsmay comprise sodium carbonate residuals of less than 100 ppm as measuredusing a high-performance liquid chromatography sodium carbonate assay.The method may further comprise adding a therapeutic agent to theaqueous solution of pure silk fibroin-based protein fragments. Themethod may further comprise adding a molecule selected from one of anantioxidant or an enzyme to the aqueous solution of pure silkfibroin-based protein fragments. The method may further comprise addinga vitamin to the aqueous solution of pure silk fibroin-based proteinfragments. The vitamin may be vitamin C or a derivative thereof. Themethod may further comprise adding an alpha hydroxy acid to the aqueoussolution of pure silk fibroin-based protein fragments. The alpha hydroxyacid may be selected from the group consisting of glycolic acid, lacticacid, tartaric acid and citric acid. The method may further compriseadding hyaluronic acid or its salt form at a concentration of about 0.5%to about 10.0% to the aqueous solution of pure silk fibroin-basedprotein fragments. The method may further comprise adding at least oneof zinc oxide or titanium dioxide.

A method for preparing an aqueous solution of pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from about 17 kDa to about 38 kDa includes the steps of: addinga silk source to a boiling (100° C.) aqueous solution of sodiumcarbonate for a treatment time of between about 30 minutes to about 60minutes so as to result in degumming; removing sericin from the solutionto produce a silk fibroin extract comprising non-detectable levels ofsericin; draining the solution from the silk fibroin extract; dissolvingthe silk fibroin extract in a solution of lithium bromide having astarting temperature upon placement of the silk fibroin extract in thelithium bromide solution that ranges from about 80° C. to about 140° C.;maintaining the solution of silk fibroin-lithium bromide in a dry ovenhaving a temperature in the range between about 60° C. to about 100° C.for a period of at least 1 hour; removing the lithium bromide from thesilk fibroin extract; and producing an aqueous solution of pure silkfibroin-based protein fragments, wherein the aqueous solution of puresilk fibroin-based protein fragments comprises lithium bromide residualsof between about 10 ppm and about 300 ppm, wherein the aqueous solutionof silk protein fragments comprises sodium carbonate residuals ofbetween about 10 ppm and about 100 ppm, wherein the aqueous solution ofpure silk fibroin-based protein fragments comprises fragments having anaverage weight average molecular weight ranging from about 17 kDa toabout 38 kDa, and wherein the aqueous solution of pure silkfibroin-based protein fragments comprises a polydispersity of betweenabout 1.5 and about 3.0. The method may further comprise drying the silkfibroin extract prior to the dissolving step. The aqueous solution ofpure silk fibroin-based protein fragments may comprise lithium bromideresiduals of less than 300 ppm as measured using a high-performanceliquid chromatography lithium bromide assay. The aqueous solution ofpure silk fibroin-based protein fragments may comprise sodium carbonateresiduals of less than 100 ppm as measured using a high-performanceliquid chromatography sodium carbonate assay. The method may furthercomprise adding a therapeutic agent to the aqueous solution of pure silkfibroin-based protein fragments. The method may further comprise addinga molecule selected from one of an antioxidant or an enzyme to theaqueous solution of pure silk fibroin-based protein fragments. Themethod may further comprise adding a vitamin to the aqueous solution ofpure silk fibroin-based protein fragments. The vitamin may be vitamin Cor a derivative thereof. The method may further comprise adding an alphahydroxy acid to the aqueous solution of pure silk fibroin-based proteinfragments. The alpha hydroxy acid may be selected from the groupconsisting of glycolic acid, lactic acid, tartaric acid and citric acid.The method may further comprise adding hyaluronic acid or its salt format a concentration of about 0.5% to about 10.0% to the aqueous solutionof pure silk fibroin-based protein fragments. The method may furthercomprise adding at least one of zinc oxide or titanium dioxide.

A method for preparing an aqueous solution of pure silk fibroin-basedprotein fragments having an average weight average molecular weightranging from about 39 kDa to about 80 kDa, includes the steps of: addinga silk source to a boiling (100° C.) aqueous solution of sodiumcarbonate for a treatment time of about 30 minutes so as to result indegumming; removing sericin from the solution to produce a silk fibroinextract comprising non-detectable levels of sericin; draining thesolution from the silk fibroin extract; dissolving the silk fibroinextract in a solution of lithium bromide having a starting temperatureupon placement of the silk fibroin extract in the lithium bromidesolution that ranges from about 80° C. to about 140° C.; maintaining thesolution of silk fibroin-lithium bromide in a dry oven having atemperature in the range between about 60° C. to about 100° C. for aperiod of at least 1 hour; removing the lithium bromide from the silkfibroin extract; and producing an aqueous solution of pure silkfibroin-based protein fragments, wherein the aqueous solution of puresilk fibroin-based protein fragments comprises lithium bromide residualsof between about 10 ppm and about 300 ppm, sodium carbonate residuals ofbetween about 10 ppm and about 100 ppm, fragments having an averageweight average molecular weight ranging from about 40 kDa to about 65kDa, and wherein the aqueous solution of pure silk fibroin-based proteinfragments comprises a polydispersity of between about 1.5 and about 3.0.The method may further comprise drying the silk fibroin extract prior tothe dissolving step. The aqueous solution of pure silk fibroin-basedprotein fragments may comprise lithium bromide residuals of less than300 ppm as measured using a high-performance liquid chromatographylithium bromide assay. The aqueous solution of pure silk fibroin-basedprotein fragments may comprise sodium carbonate residuals of less than100 ppm as measured using a high-performance liquid chromatographysodium carbonate assay. The method may further comprise adding atherapeutic agent to the aqueous solution of pure silk fibroin-basedprotein fragments. The method may further comprise adding a moleculeselected from one of an antioxidant or an enzyme to the aqueous solutionof pure silk fibroin-based protein fragments. The method may furthercomprise adding a vitamin to the aqueous solution of pure silkfibroin-based protein fragments. The vitamin may be vitamin C or aderivative thereof. The method may further comprise adding an alphahydroxy acid to the aqueous solution of pure silk fibroin-based proteinfragments. The alpha hydroxy acid may be selected from the groupconsisting of glycolic acid, lactic acid, tartaric acid and citric acid.The method may further comprise adding hyaluronic acid or its salt format a concentration of about 0.5% to about 10.0% to the aqueous solutionof pure silk fibroin-based protein fragments. The method may furthercomprise adding at least one of zinc oxide or titanium dioxide.

Example 8. Characterization of Silk Coatings on Polyester

A summary of the results from studies of silk coatings on polyester aregiven in Table 29 and Table 30. The results shown in FIG. 93 and FIG. 94illustrate that the accumulative one way transport index and OMMCperformance is maintained even at 60 wash cycles. Additional testresults are shown in FIG. 95 to FIG. 102 . The antimicrobial performanceof the silk coated polyester fabrics are maintained to 25 to 50 washingcycles, as shown in FIG. 103 to FIG. 104 . The results illustrate thesurprising improvement in moisture management properties, as well as thesurprising result that the improved properties survive many wash cycles.

TABLE 29 Test results for semifinished polyester with 1% silk solutioncoating. Testing Results: Semifinished polyester with 1% silk solutioncoating Top Bottom Overall Wetting Wetting Top Bottom Max Max Top BottomAccumulative Moisture Time Time Absorption Absorption Wetted WettedSpreading Spreading One-Way Management Number of Top Bottom Rate RateRadius Radius Speed Speed Transport Capability Washes Raw Data: (sec)(sec) (%/sec) (%/sec) (mm) (mm) (mm/sec) (mm/sec) index(%) OMMC  0Cycles Mean 5.63 3.95 7.24 28.73 5 5 0.90 1.22 133.26 0.27 10 Cycles S.Deviation 1.20 0.38 1.46 8.62 0 0 0.20 0.12 34.81 0.06 25 Cycles CV 0.210.10 0.20 0.30 0 0 0.22 0.09 0.26 0.21 50 Cycles Mean 23.87 7.96 4.828.55 5 5 0.46 0.68 144.84 0.22 S. Deviation 31.51 3.30 0.84 2.94 0 00.28 0.23 27.71 0.03 CV 1.32 0.41 0.17 0.34 0 0 0.61 0.33 0.19 0.14 Mean6.09 4.59 7.36 17.22 5 5 0.83 1.05 124.05 0.22 S. Deviation 1.61 0.442.98 3.28 0 0 0.17 0.09 11.70 0.02 CV 0.26 0.10 0.40 0.19 0 0 0.20 0.090.09 0.09 Mean 25.20 11.64 6.84 7.80 5 5 0.39 0.53 58.81 0.13 S.Deviation 28.06 6.36 3.38 5.70 0 0 0.30 0.27 26.56 0.03 CV 1.11 0.550.49 0.73 0 0 0.77 0.51 0.45 0.25

TABLE 30 Test results for wicking finished polyester without silkcoating. Testing Results: Wicking Finished Polyester Top Bottom OverallWetting Wetting Top Bottom Max Max Top Bottom Accumulative Moisture TimeTime Absorption Absorption Wetted Wetted Spreading Spreading One-WayManagement Number of Top Bottom Rate Rate Radius Radius Speed SpeedTransport Capability Washes Raw Data: (sec) (sec) (%/sec) (%/sec) (mm)(mm) (mm/sec) (mm/sec) index(%) OMMC  0 Cycles Mean 3.46 3.48 37.3056.90 5 5 1.37 1.36 62.37 0.29 S .Deviation 0.07 0.04 12.89 10.24 0 00.02 0.02 9.74 0.03 CV 0.02 0.01 0.35 0.18 0 0 0.02 0.01 0.16 0.12 25Cycles Mean 6.69 6.71 7.23 6.89 5 5 0.75 0.76 30.40 0.09 S. Deviation1.48 1.92 1.27 2.74 0 0 0.13 0.19 16.22 0.02 CV 0.22 0.29 0.18 0.40 0 00.17 0.25 0.53 0.20 50 Cycles Mean 11.27 8.46 6.70 9.35 5 5 0.54 0.6531.21 0.09 S. Deviation 6.57 3.53 1.45 5.21 0 0 0.23 0.25 18.26 0.03 CV0.58 0.42 0.22 0.56 0 0 0.44 0.38 0.59 0.30

Example 9. Characterization of Silk Coatings on Polyester Fabrics

Scanning electron microscopy (SEM) analysis was performed using aHitachi S-4800 field-emission SEM (FE-SEM) operated at 2 kV acceleratingvoltage. Pieces from each sample were sectioned using a razor blade andplaced on carbon adhesive tape mounted on aluminum SEM stubs. A coatingof iridium approximately 2 nm thick was applied via sputter depositionin order to minimize the buildup of surface charge.

The samples used in the SEM study are described in Table 31. SEMmicrographs for fabric samples are shown in FIG. 105 to FIG. 167 .

TABLE 31 Fabric samples tested by scanning electron microscopy andoptical profilometry. Silk solution for coating/treatment Silkcoating/treatment (average molecular method using silk Sample ID Fabricweight, Da) fibroin solution (sfs) FAB-10-SPRAY-B Semi-finished UA41.576 spray with 1% sfs R20904012 FAB-01-SPRAY-B Semi-finished UA41.576 spray with 0.1% sfs R20904012 FAB-10-STEN-B Semi-finished UA41.576 stencil spray with 1% sfs R20904012 FAB-10-BATH-B Semi-finishedUA 41.576 bath with 1% sfs R20904012 FAB-01-BATH-B Semi-finished UA41.576 bath with 0.1% sfs R20904012 FAB-01-SPRAY-C Semi-finished UA10.939 spray with 0.1% sfs R20904012 FAB-01-STEN-C Semi-finished UA10.939 stencil spray with 0.1% sfs R20904012 FAB-10-BATH-C Semi-finishedUA 10.939 bath with 1% sfs R20904012

The fabric SEM results show that the silk solution has very clearly beendeposited along and between individual polyester fibers. The use of 0.1%silk solution results in less coating than 1.0% silk solution. The useof a bath for 0.1% silk solution, with an average molecular weight of 41kDa, results in uniform coating along fibers with large, smoothfeatures. The use of a spray with a 0.1% silk solution, with an averagemolecular weight of 41 kDa, in coating along fibers as well asconnected, webbed coating between fibers. The use of a spray for 0.1%silk solution, with an average molecular weight of 11 kDa, results inuniform coating along fibers with small, spotted/ribbed features. Theuse of a stencil for 0.1% silk solution, with an average molecularweight of 11 kDa, results in coating along fibers that has clear edgesand delineation between coated and non coated sides. The use of a bathfor 1.0% silk solution, with an average molecular weight of 41 kDa,results in thick coating along fibers as well as thick connected, webbedcoating between fibers. The use of a bath for 1.0% silk solution, withan average molecular weight of 11 kDa, results in coating along allsides of individual fibers. Coating appears uniform on surface with manysingle point extrusions. The use of a spray for 1.0% silk solution, withan average molecular weight of 41 kDa, results in coating along fibersas well as connected, webbed coating between fibers, which was thickerthan that observed using 0.1% silk solution. The use of a stencil for1.0% silk solution, with an average molecular weight of 41 kDa, resultsin coating along fibers and between fibers, and the coating appears wellorganized.

The SEM results demonstrate that the silk coating has been applied as aneven, thin, uniform coating to the fibers of the fabric. Thisillustrates the surprising result that the silk coating was applied tothe fibers without the use of any additives or cross-linking, using awater based delivery system.

Example 10. Characterization of Silk Coatings on Polyester Films

The film samples are described in Table 32. The SEM images from thesefilms are shown in FIG. 168 to FIG. 237 .

TABLE 32 Film samples tested by scanning electron microscopy and opticalprofilometry. Silk solution for Polyester coating/treatment Silkcoating/treatment substrate (average molecular method using silk Sampleidentifier material weight, Da) fibroin solution (sfs)FIL-10-SPRAY-B-01MYL 0.01 Mylar 41.576 spray with 1% sfsFIL-01-SPRAY-B-01MYL 0.01 Mylar 41.576 spray with 0.1% sfsFIL-01-SPRAY-B-007MEL  0.007 Melinex 41.576 spray with 0.1% sfsFIL-01-SPRAY-C-01MYL 0.01 Mylar 10.939 spray with 0.1% sfsFIL-01-STEN-B-01MYL 0.01 Mylar 41.576 stencil spray with 0.1% sfsFIL-01-STEN-C-01MYL 0.01 Mylar 10.939 stencil spray with 0.1% sfsFIL-10-BATH-B-01MYL 0.01 Mylar 41.576 bath with 1% sfsFIL-10-BATH-B-007MEL  0.007 Melinex 41.576 bath with 1% sfsFIL-10-BATH-C-01MYL 0.01 Mylar 10.939 bath with 1% sfsFIL-01-BATH-B-01MYL 0.01 Mylar 41.576 bath with 0.1% sfs

The results show that the silk coatings are applied uniformly. Little tono variation is observed in the characteristics or topology of thecoated polyester films. Surprisingly, the coating is even, uniform, andthin. Furthermore, surprising, the silk coated the fibers without anyadditives or cross-linking using a water-based system.

Optical profiling was carried out using a Zygo New View 6200 opticalprofilometer. Two locations on each sample were randomly selected andmeasured with 10× magnification. The results are shown in FIG. 241 toFIG. 264 . The results indicate that the silk-coated samples have ahomogeneous deposition of silk fibroin. Surface roughness featuresobserved in the control are visible after silk coating on Mylar films,which is consistent with the presence of a relatively thin silk filmthat is forming a conformal coating on Mylar. The results substantiatethe uniformity of the coating, and demonstrate that silk can bestenciled into discrete locations.

Contact profilometry was performed and the cross-sectioned samples wereexamined by SEM. Results are shown in FIG. 265 to FIG. 268 . For sampleFIL-10-SPRAY-B-10MYL, the thickness ranged from approximately 260 nm to850 nm in 4 locations analyzed. For sample FIL-10-BATH-B-01MYL, thecoating ranged from approximately 140 nm to 400 nm in 4 locations. SEMimages from cross-sections show similar trends, with one location onsample FIL-10-SPRAY-B-10MYL having a cross-section that measuresapproximately 500 nm and one on FIL-10-BATH-B-01MYL measuringapproximately 180 nm.

Example 11. Preparation of Silk Fibroin Solutions with Higher MolecularWeights

The preparation of silk fibroin solutions with higher molecular weightsis given in Table 33.

TABLE 33 Preparation and properties of silk fibroin solutions. Averageweight average Extraction molecular Time Extraction LiBr Temp Oven/Sol’nweight Average Sample Name (mins) Temp (° C.) (° C.) Temp (kDa)polydispersity Group A TFF 60 100 100 100° C. oven 34.7 2.94 Group A DIS60 100 100 100° C. oven 44.7 3.17 Group B TFF 60 100 100 100° C. sol’n41.6 3.07 Group B DIS 60 100 100 100° C. sol’n 44.0 3.12 Group C TFF 60100 140 140° C. sol’n 10.9 3.19 Group C DIS 60 100 140 140° C. sol’nGroup D DIS 30  90  60  60° C. sol’n 129.7 2.56 Group D FIL 30  90  60 60° C. sol’n 144.2 2.73 Group E DIS 15 100 RT  60° C. sol’n 108.8 2.78Group E FIL 15 100 RT  60° C. sol’n 94.8 2.62

Example 12. Silk Coatings on Natural Fabrics

The coating of natural fabric with silk fibroin solutions and theresulting properties are illustrated in Table 34, Table 35, FIG. 269 ,and FIG. 270 . The results demonstrate that silk fibroin solutions cancoat cotton-Lyrca natural fabrics including LUON and POWER LUXTREME.

TABLE 34 Silk fibroin coated fabrics. Legend Fabric 15072201 PowerLuxtreme RT1211362 15072202 Luon RT20602020 15072301 Power LuxtremeRT1211362 (15072201) 1% silk solution spray coating 15072302 LuonRT20602020 (15072202) 1% silk solution spray coating 15072303 PowerLuxtreme RT1211362 (15072201) 0.1% silk solution spray coating 15072304Luon RT20602020 (15072202) 0.1% silk solution spray coating 15072305Power Luxtreme RT1211362 (15072201) 1% silk solution stencil coating15072306 Luon RT20602020 (15072202) 1% silk solution stencil coating15072307 Power Luxtreme RT1211362 (15072201) 0.1% silk solution stencilcoating 15072308 Luon RT20602020 (15072202) 0.1% silk solution stencilcoating 15072309 Power Luxtreme RT1211362 (15072201) 1% silk solutionbath coating 15072310 Luon RT20602020 (15072202) 1% silk solution bathcoating 15072311 Power Luxtreme RT1211362 (15072201) 0.1% silk solutionbath coating 15072312 Luon RT20602020 (15072202) 0.1% silk solution bathcoating

TABLE 35 Test results for silk fibroin coated fabrics. Top BottomOverall Wetting Wetting Top Bottom Max Max Top Bottom AccumulativeMositure Time Time Absorption Absorption Wetted Wetted SpreadingSpreading One-Way Management Raw Top Bottom Rate Rate Radius RadiusSpeed Speed Transport Capability Data: (sec) (sec) (%/sec) (%/sec) (m m)(mm) (mm/sec) (mm/sec) index (%) OMMC 15072201 Mean 64.3786 3.40728.8123 8.60494 5 5 0.15038 1.41686 151.65248 0.25898 15072202 Mean25.1786 28.1922 5.4636 6.195 5 5 0.218 0.4244 80.9572 0.1529 15072301Mean 16.7172 12.2804 21.9859 33.6196 5 5 0.4304 0.4906 143.6659 0.280815072302 Mean 25.8898 41.5026 6.16512 8.70282 5 5 0.23336 0.179144.06124 0.10704 15072303 Mean 42.152 4.7268 7.9114 19.3725 4 5 0.32611.364 370.2757 0.5297 15072304 Mean 78.4746 34.3138 5.01486 6.63212 5 50.0661 0.38728 94.97976 0.16848 15072305 Mean 36.1954 17.2038 6.271586.25526 5 5 0.18872 0.89046 139.73478 0.23052 15072306 Mean 78.474634.3138 5.01486 6.63212 5 5 0.0661 0.38728 94.97976 0.16848 15072307Mean 36.195 17.2038 6.2716 6.2553 5 5 0.1887 0.8905 139.7348 0.230515072308 Mean 57.335 25.7588 5.6432 6.4437 5 5 0.1274 0.6389 117.35730.1995 15072309 Mean 54.1384 9.2662 4.01594 9.11064 5 5 0.09398 0.85306267.0755 0.36724 15072310 Mean 28.4544 13.6658 6.8844 7.8956 5 5 0.30590.5111 104.5035 0.1794 15072311 Mean 5.1292 4.4738 8.8047 13.0277 5 50.9486 1.1702 246.6729 0.3597 15072312 Mean 6.8516 9.4722 11.068411.7268 5 5 0.7394 0.5794 73.4005 0.1461

All patents, patent applications, and published references cited hereinare hereby incorporated by reference in their entirety. While themethods of the present disclosure have been described in connection withthe specific embodiments thereof, it will be understood that it iscapable of further modification. Further, this application is intendedto cover any variations, uses, or adaptations of the methods of thepresent disclosure, including such departures from the presentdisclosure as come within known or customary practice in the art towhich the methods of the present disclosure pertain.

1. An article comprising a fiber or yarn having a coating, wherein thecoating comprises silk based proteins or fragments thereof having aweight average molecular weight range of about 5 kDa to about 144 kDa.2. The article of claim 1, wherein the article is a fabric.
 3. Thearticle of claim 1, wherein the silk based proteins or fragments thereofcomprise silk fibroin-based proteins or protein fragments having about0.01% (w/w) to about 10% (w/w) sericin.
 4. The article of claim 1,wherein the silk based proteins or fragments thereof are selected fromthe group consisting of natural silk based proteins or fragmentsthereof, recombinant silk based proteins or fragments thereof, andcombinations thereof.
 5. The article of claim 4, wherein the silk basedproteins or fragments thereof are natural silk based proteins orfragments thereof that are selected from the group consisting of spidersilk based proteins or fragments thereof, silkworm silk based proteinsor fragments thereof, and combinations thereof.
 6. The article of claim5, wherein the natural silk based proteins or fragments are silkwormsilk based proteins or fragments thereof, and the silkworm silk basedproteins or fragments thereof is Bombyx mori silk based proteins orfragments thereof.
 7. The article of claim 1, wherein the silk basedproteins or fragments comprise silk and a copolymer.
 8. The article ofclaim 1, wherein the silk based proteins or protein fragments thereofhave an average weight average molecular weight range selected from thegroup consisting of about 5 to about 10 kDa, about 6 kDa to about 16kDa, about 17 kDa to about 38 kDa, about 39 kDa to about 80 kDa, about60 to about 100 kDa, and about 80 kDa to about 144 kDa, wherein the silkbased proteins or fragments thereof have a polydispersity of betweenabout 1.5 and about 3.0, and wherein the proteins or protein fragments,prior to coating the fabric, do not spontaneously or gradually gelateand do not visibly change in color or turbidity when in a solution forat least 10 days.
 9. The article of claim 1, wherein the fiber or yarnis selected from the group consisting of natural fiber or yarn,synthetic fiber or yarn, or combinations thereof.
 10. The article ofclaim 9, wherein the fiber or yarn is natural fiber or yarn selectedfrom the group consisting of cotton, alpaca fleece, alpaca wool, lamafleece, lama wool, cotton, cashmere, sheep fleece, sheep wool, andcombinations thereof.
 11. The article of claim 9, wherein the fiber oryarn is synthetic fiber or yarn selected from the group consisting ofpolyester, nylon, polyester-polyurethane copolymer, and combinationsthereof.
 12. The article of claim 2, wherein the fabric exhibits animproved property, wherein the improved property is an accumulativeone-way moisture transport index selected from the group consisting ofgreater than 40%, greater than 60%, greater than 80%, greater than 100%,greater than 120%, greater than 140%, greater than 160%, and greaterthan 180%.
 13. The article of claim 2, wherein the fabric exhibits animproved property, wherein the improved property is an accumulative oneway transport capability increase relative to uncoated fabric selectedfrom the group consisting of 1.2 fold, 1.5 fold, 2.0 fold, and 3.0 fold.14. The article of claim 2, wherein the fabric exhibits an improvedproperty, wherein the improved property is an overall moisturemanagement capability selected from the group consisting of greater than0.05, greater than 0.10, greater than 0.15, greater than 0.20, greaterthan 0.25, greater than 0.30, greater than 0.35, greater than 0.40,greater than 0.50, greater than 0.60, greater than 0.70, and greaterthan 0.80.
 15. The article of claim 14, wherein the improved property isdetermined after a period of machine washing cycles selected from thegroup consisting of 5 cycles, 10 cycles, 25 cycles, and 50 cycles. 16.The article of claim 2, wherein the fabric exhibits substantially noincrease in microbial growth after a number of machine washing cyclesselected from the group consisting of 5 cycles, 10 cycles, 25 cycles,and 50 cycles.
 17. The article of claim 16, wherein the microbial growthis microbial growth of a microbe selected from the group consisting ofStaphylococcus aureus, Klebisiella pneumoniae, and combinations thereof.18. The article of claim 17, wherein the microbial growth is reduced bya percentage selected from the group consisting of 50%, 100%, 500%,1000%, 2000%, and 3000% compared to an uncoated fabric.
 19. The articleof claim 2, wherein the coating is applied to the fabric at the fiberlevel prior to forming the fabric.
 20. The article of claim 2, whereinthe coating is applied to the fabric at the fabric level. 21-30.(canceled)